Patent Publication Number: US-2022233682-A1

Title: Vaccine compositions for the treatment of coronavirus

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     This application is a continuation-in-part of U.S. application Ser. No. 17/218,148, filed Mar. 30, 2021 which claims the benefit of U.S. Provisional Application No. 63/002,237, filed Mar. 30, 2020, and of U.S. Provisional Application No. 63/070,150, filed Aug. 25, 2020, the contents of each of which are hereby incorporated herein in their entireties. 
    
    
     SEQUENCE LISTING 
     The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Mar. 31, 2022, is named 2007801-0148_SL.txt and is 205,747 bytes in size. 
     FIELD OF THE INVENTION 
     This invention is in the field of vaccines, in particular virus like particle vaccines for coronavirus. 
     BACKGROUND 
     Coronaviruses are spherical, enveloped viruses, ranging from 160-180 nm in diameter and containing a positive-stranded RNA genome. With their genome of approximately 30,000 bases, they are considered the largest of the known RNA viruses. Like influenza viruses they have the ability to genetically recombine with other members of the coronavirus family. Coronaviruses fall into four major genera. Coronaviruses are believed to be the causative agents of several severe diseases in many animals, for example, infectious bronchitis virus, feline infectious peritonitis virus and transmissible gastroenteritis virus. Coronaviruses also cause a range of illnesses in humans from the common cold to severe respiratory infections. Four human coronaviruses, HCoV-0C43, HCoV-HKU1 (betacoronaviruses), and HCoV-NL63, HCoV-229E (alphacoronaviruses), contribute to 15%-30% of common colds (Fung et al (2019) Annu. Rev. Microbiol. 73:2-529-557). In recent years, beta-coronaviruses have been responsible for three significant outbreaks of disease in humans. 
     In the early 2000s, a beta coronavirus known as SARS-CoV caused an outbreak of respiratory disease referred to as severe acute respiratory syndrome (SARS). The main symptoms included fever, dry cough, headache, shortness of breath and difficulty of breathing. Many of those infected developed viral pneumonia resulting in infection of the lower respiratory tract. SARS is highly contagious, and is spread by droplets caused by coughing or sneezing or through other methods such as fecal contamination. SARS was fatal in around 9.14% of all cases. The global outbreak of SARS was contained in July 2003 and there have been no reported cases since 2004 (Peeri et al Int. J. Epi, Feb. 10, 2020). 
     In 2012, another novel coronavirus emerged in Saudi Arabia which is now known as Middle East Respiratory Syndrome coronavirus (MERS-CoV). MERS-CoV is also beta coronavirus. Subsequent cases of MERS-CoV infection were reported and the outbreak spread to 27 countries in the Middle East, Europe, Asia and North America. Infection with MERS-CoV presented as a severe acute respiratory illness with symptoms of fever, cough, and shortness of breath. About 34% of reported cases of MERS-CoV infection resulted in death. Only a small number of reported cases involved subjects with mild respiratory illness. 
     In late 2019, a respiratory infection appeared in Wuhan, China which was quickly identified as caused by a novel coronavirus strain called SARS-CoV-2. The infection, known as COVID-19 is highly infectious and causes severe pneumonia, particularly in elderly patients. Mortality rates vary significantly by country, with estimates ranging from 13.7% in Italy to 1.9% in Japan. As of March 2021, the fatality rate in the United States was approximately 1.8% (Johns Hopkins Coronavirus Research Centre, Update as of Mar. 30, 2021). COVID-19 quickly spread throughout the world resulting in a significant threat to human health and a massive slowdown in economic activity. As of Feb. 1, 2021, more than 100 million people had contracted COVID-19, and over 2 million had died. 
     In late 2020, several vaccines against COVID-19 were approved for emergency use. These vaccines target a protein on the surface of SARS-CoV-2 known as the spike protein and utilized novel platforms, sometimes for the first time for human use. These vaccines were shown to be highly effective in clinical trials, but distribution has been slow in many parts of the world due to manufacturing challenges and, in some cases, the requirement for storage at ultra-low temperatures. Furthermore, while several new vaccines have proven to be safe, some have been associated with rare but deadly side effects that have restricted their use in certain countries. 
     During the second half of 2020, variants of SARS-CoV-2 emerged which cause COVID-19 disease, and those variants which had an impact on transmissibility, severity of disease and/or immunity were designated Variants of Concern (VoCs). Three VoCs rapidly became dominant during this period in the countries where they were first detected, B.1.1.7 first identified in the UK (also known as the Alpha variant), 501Y.V2 or B.1351 first identified in South Africa (also known as the Beta variant), and P.1 first identified in Brazil (also known as the Gamma variant). More recently, new VoCs emerged in India (B.1617 also known as the Delta variant) and in South Africa (B.1.1.529 also known as the Omicron variant). These VoCs have proven to be highly infectious due to increased binding affinity of the viral receptor-binding domain (RBD) to the receptor known as angiotensin-converting enzyme 2 (ACE2). Each of the VoCs are characterized by a number of shared mutations expressed on the spike protein, primarily located in the RBD and N-terminal domain (NTD), that serve to increase transmission and/or enhance escape from neutralizing antibodies acquired by vaccination or prior natural SARS-CoV-2 infection. The rapid spread of the new VoCs, and the possible emergence of new variants has raised significant concerns regarding reinfection and the effectiveness of the recently approved vaccines, all of which were developed against the original strain of SARS-CoV-2. 
     As a result, there is an urgent need to develop new vaccines which induce strong immunity against SARS-CoV-2 while being safe and easy to store and distribute. Furthermore, there is an urgent need to ensure that vaccines against SARS-CoV-2 provide broad immunity so as to protect patients against mutated forms of the virus including VoCs. 
     Accordingly, a need exists for a vaccine against human coronaviruses which provides broad immunity against coronavirus antigens. 
     SUMMARY 
     The present disclosure provides methods and compositions useful for prophylaxis of infection cause by human coronaviruses. More particularly, the present disclosure provides methods for production of, and compositions comprising, virus like particles (VLPs) expressing antigens from human coronaviruses which are useful for prevention, treatment, and/or diagnosis of infections caused by coronaviruses. 
     The present disclosure provides virus-like particles which comprise one or more Moloney Murine leukemia virus (MMLV) core proteins and are surrounded by a lipid bilayer membrane. The VLPs include one or more envelope polypeptides from human coronaviruses (e.g., one or more coronavirus polypeptide epitopes) that play a role in induction of virus-neutralizing antibodies. 
     In some embodiments, the present disclosure provides VLPs having an envelope that comprises a wild type human coronavirus envelope glycoprotein. In some embodiments, the polypeptide is from SARS-CoV. In some embodiments, the polypeptide is from MERS-CoV. In some embodiments, the polypeptide is from SARS-CoV-2. The polypeptides from SARS-CoV-2 can be from the ancestral strain, first identified in Wuhan China, or from a variant of the ancestral strain. In some embodiments, the VLPs include polypeptides from more than one of SARS-CoV, MERS-CoV and SARS-CoV-2. In some embodiments, the VLPs include polypeptides from all three of SARS-CoV, MERS-CoV and SARS-CoV-2. 
     In some embodiments, the present disclosure provides VLPs having an envelope that comprises a modified human coronavirus envelope glycoprotein. In an embodiment, the present disclosure encompasses production of VLPs having envelopes that include a coronavirus polypeptide in a premature conformation. In a specific embodiment, the modified envelope glycoprotein lacks a furin cleavage site. In some embodiments, the polypeptide lacking a furin cleavage site is from SARS-CoV. In some embodiments, the polypeptide lacking a furin cleavage site is from MERS-CoV. In some embodiments, the polypeptide lacking a furin cleavage site is from SARS-CoV-2. In some embodiments, the VLPs include polypeptides from more than one of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides lack a furin cleavage site. In some embodiments, the VLPs include polypeptides from all three of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides lack a furin cleavage site. 
     In another embodiment, the present disclosure encompasses production of VLPs having envelopes that include a coronavirus polypeptide having a modified amino acid sequence. In a specific embodiment, the modified envelope glycoprotein has a lysine and valine residue substituted for proline residues. In some embodiments, the polypeptide having a proline substitution is from SARS-CoV. In some embodiments, the polypeptide having a proline substitution is from MERS-CoV. In some embodiments, the polypeptide having a proline substitution is from SARS-CoV-2. In some embodiments, the VLPs include polypeptides from more than one of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides have a proline substitution. In some embodiments, the VLPs include polypeptides from all three of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides have a proline substitution. 
     In another embodiment, the present disclosure encompasses production of VLPs having envelopes that include a coronavirus polypeptide having a modified amino acid sequence and a premature conformation. In a specific embodiment, the modified envelope glycoprotein has a lysine and valine residue substituted for proline residues and lack a furin cleavage site. In some embodiments, the polypeptide having a proline substitution and lacking a furin cleavage site is from SARS-CoV. In some embodiments, the polypeptide having a proline substitution and lacking a furin cleavage site is from MERS-CoV. In some embodiments, the polypeptide having a proline substitution and lacking a furin cleavage site is from SARS-CoV-2. In some embodiments, the VLPs include polypeptides from more than one of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides have a proline substitution and lack a furin cleavage site. In some embodiments, the VLPs include polypeptides from all three of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides have a proline substitution and lack a furin cleavage site. 
     In a further embodiment, the modified envelope glycoprotein has been modified such that the transmembrane domain is replaced with the transmembrane domain of another virus. In a particularly preferred embodiment, the VLP has a modified envelope glycoprotein comprising an isolated coronavirus S protein, the transmembrane domain and cytoplasmic tail of which protein have been replaced with the transmembrane domain and cytoplasmic tail from vesicular stomatitis virus (VSV). In some embodiments, the polypeptide having a transmembrane domain and cytoplasmic tail from VSV is from SARS-CoV. In some embodiments, the polypeptide having a transmembrane domain and cytoplasmic tail from VSV is from MERS-CoV. In some embodiments, the polypeptide having a transmembrane domain and cytoplasmic tail from VSV is from SARS-CoV-2. In some embodiments, the VLPs include polypeptides from more than one of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides have a transmembrane domain and cytoplasmic tail from VSV. In some embodiments, the VLPs include polypeptides from all three of SARS-CoV, MERS-CoV and SARS-CoV-2, wherein the polypeptides have a transmembrane domain and cytoplasmic tail from VSV. In some embodiments, the VLPS include one or more polypeptides from SARS-CoV, MERS-CoV and SARS-CoV-2, one or more of which have been modified as described herein and which have a transmembrane domain and cytoplasmic tail from VSV. 
     In a preferred embodiment, the present disclosure encompasses production of a VLP having an envelope that includes a SARS-CoV-2 spike polypeptide having a modified amino acid sequence and a premature conformation. The modified envelope glycoprotein has a lysine and valine residue substituted for proline residues and it lacks a furin cleavage site. Furthermore, the modified spike glycoprotein has been further modified such that the transmembrane domain and cytoplasmic tail have been replaced with the transmembrane domain and cytoplasmic tail from vesicular stomatitis virus (VSV). 
     The present disclosure further provides bivalent and trivalent VLPs comprising one or more modified human coronavirus envelope proteins and one or more wild type human coronavirus proteins. 
     The present disclosure further provides VLPs having an envelope that includes a SARS-CoV-2 spike polypeptide, wild type or modified, from a variant of the ancestral SARS-CoV-2 strain which comprises a heterologous booster vaccine. In a preferred embodiment, the VLP includes a modified SARS-CoV-2 spike polypeptide from the Beta variant. In a particularly preferred embodiment, the SARS-CoV-2 spike polypeptide from the Beta variant has a lysine and valine residue substituted for proline residues, it lacks a furin cleavage site and has been further modified such that the transmembrane domain and cytoplasmic tail have been replaced with the transmembrane domain and cytoplasmic tail from vesicular stomatitis virus (VSV). 
     Other features, objects, and advantages of the present invention are apparent in the detailed description that follows. It should be understood, however, that the detailed description, while indicating embodiments of the present invention, is given by way of illustration only, not limitation. Various changes and modifications within the scope of the invention will become apparent to those skilled in the art from the detailed description. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The drawings are for illustration purposes only, not for limitation. 
         FIG. 1  is a diagram illustrating the structure of the SARS-CoV-2 envelope. 
         FIG. 2  the S1/S2 domains from SARS-CoV, SARS CoV-2 and MERS-CoV. 
         FIG. 3  is a diagram illustrating exemplary alternative COVID-S constructs. 
         FIG. 3  discloses “RRAR” as SEQ ID NO:46 and “GSAS” as SEQ ID NO:47. 
       
         
           
             
                 
               
                 
                     
                 
                 
                   Listing of Sequences 
                 
                 
                     
                 
               
              
                 
                     
                 
              
             
             
                 
              
                 
                   The following is a list of sequences referred to herein: 
                 
                 
                   SEQ ID NO: 1 is an MMLV-Gag Amino Acid Sequence 
                 
                 
                   MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTFNVGWPRDGTFNR 
                 
                 
                   DLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKPPPPLPPSAPSLPLEP 
                 
                 
                   PRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTEDPPPYRDPRPPPSDRDGNGGEA 
                 
                 
                   TPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLRAGGNGQLQYWPFSSSDLYNWKNN 
                 
                 
                   NPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGTLLTGEEKQRVLLEARKAVRGDDGRP 
                 
                 
                   TQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYRQLLLAGLQNAGRSPTNLAKVKGITQ 
                 
                 
                   GPNESPSAFLERLKEAYRRYTPYDPEDPGQETNVSMSFIWQSAPDIGRKLERLEDLKNKTL 
                 
                 
                   GDLVREAEKIFNKRETPEEREERIRRETEEKEERRRTEDEQKEKERDRRRHREMSKLLAT 
                 
                 
                   VVSGQKQDRQGGERRRSQLDRDQCAYCKEKGHWAKDCPKKPRGPRGPRPQTSLLTLDD 
                 
                 
                     
                 
                 
                   SEQ ID NO: 2 is MMLV-Gag Nucleotide Sequence 
                 
                 
                   ATGGGCCAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCG 
                 
                 
                   AGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCT 
                 
                 
                   GCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACC 
                 
                 
                   GAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCC 
                 
                 
                   AGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGG 
                 
                 
                   GTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCT 
                 
                 
                   CCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTC 
                 
                 
                   CTTCTCTAGGCGCCAAACCTAAACCTCAAGTTCTTTCTGACAGTGGGGGGCCGCTCAT 
                 
                 
                   CGACCTACTTACAGAAGACCCCCCGCCTTATAGGGACCCAAGACCACCCCCTTCCGA 
                 
                 
                   CAGGGACGGAAATGGTGGAGAAGCGACCCCTGCGGGAGAGGCACCGGACCCCTCCC 
                 
                 
                   CAATGGCATCTCGCCTACGTGGGAGACGGGAGCCCCCTGTGGCCGACTCCACTACCT 
                 
                 
                   CGCAGGCATTCCCCCTCCGCGCAGGAGGAAACGGACAGCTTCAATACTGGCCGTTCT 
                 
                 
                   CCTCTTCTGACCTTTACAACTGGAAAAATAATAACCCTTCTTTTTCTGAAGATCCAGG 
                 
                 
                   TAAACTGACAGCTCTGATCGAGTCTGTTCTCATCACCCATCAGCCCACCTGGGACGAC 
                 
                 
                   TGTCAGCAGCTGTTGGGGACTCTGCTGACCGGAGAAGAAAAACAACGGGTGCTCTTA 
                 
                 
                   GAGGCTAGAAAGGCGGTGCGGGGCGATGATGGGCGCCCCACTCAACTGCCCAATGA 
                 
                 
                   AGTCGATGCCGCTTTTCCCCTCGAGCGCCCAGACTGGGATTACACCACCCAGGCAGG 
                 
                 
                   TAGGAACCACCTAGTCCACTATCGCCAGTTGCTCCTAGCGGGTCTCCAAAACGCGGG 
                 
                 
                   CAGAAGCCCCACCAATTTGGCCAAGGTAAAAGGAATAACACAAGGGCCCAATGAGT 
                 
                 
                   CTCCCTCGGCCTTCCTAGAGAGACTTAAGGAAGCCTATCGCAGGTACACTCCTTATGA 
                 
                 
                   CCCTGAGGACCCAGGGCAAGAAACTAATGTGTCTATGTCTTTCATTTGGCAGTCTGCC 
                 
                 
                   CCAGACATTGGGAGAAAGTTAGAGAGGTTAGAAGATTTAAAAAACAAGACGCTTGG 
                 
                 
                   AGATTTGGTTAGAGAGGCAGAAAAGATCTTTAATAAACGAGAAACCCCGGAAGAAA 
                 
                 
                   GAGAGGAACGTATCAGGAGAGAAACAGAGGAAAAAGAAGAACGCCGTAGGACAGA 
                 
                 
                   GGATGAGCAGAAAGAGAAAGAAAGAGATCGTAGGAGACATAGAGAGATGAGCAAG 
                 
                 
                   CTATTGGCCACTGTCGTTAGTGGACAGAAACAGGATAGACAGGGAGGAGAACGAAG 
                 
                 
                   GAGGTCCCAACTCGATCGCGACCAGTGTGCCTACTGCAAAGAAAAGGGGCACTGGGC 
                 
                 
                   TAAAGATTGTCCCAAGAAACCACGAGGACCTCGGGGACCAAGACCCCAGACCTCCCT 
                 
                 
                   CCTGACCCTAGATGAC 
                 
                 
                     
                 
                 
                   SEQ ID NO: 3 is a Codon Optimized MMLV-Gag Nucleotide Sequence 
                 
                 
                   ATGGGACAGACCGTCACAACACCCCTGAGCCTGACCCTGGGACATTGGAAAGACGTG 
                 
                 
                   GAGAGGATCGCACATAACCAGAGCGTGGACGTGAAGAAACGGAGATGGGTCACATT 
                 
                 
                   CTGCAGTGCTGAGTGGCCAACTTTTAATGTGGGATGGCCCCGAGACGGCACTTTCAA 
                 
                 
                   CAGGGATCTGATCACCCAGGTGAAGATCAAGGTCTTTAGCCCAGGACCTCACGGACA 
                 
                 
                   TCCAGACCAGGTGCCTTATATCGTCACCTGGGAGGCACTGGCCTTCGATCCCCCTCCA 
                 
                 
                   TGGGTGAAGCCATTTGTCCACCCAAAACCACCTCCACCACTGCCTCCAAGTGCCCCTT 
                 
                 
                   CACTGCCACTGGAACCACCCCGGAGCACACCACCCCGCAGCTCCCTGTATCCTGCTCT 
                 
                 
                   GACTCCATCTCTGGGCGCAAAGCCAAAACCACAGGTGCTGAGCGACTCCGGAGGACC 
                 
                 
                   ACTGATTGACCTGCTGACAGAGGACCCCCCACCATACCGAGATCCTCGGCCTCCACC 
                 
                 
                   AAGCGACCGCGATGGAAATGGAGGAGAGGCTACTCCTGCCGGCGAAGCCCCTGACC 
                 
                 
                   CATCTCCAATGGCTAGTAGGCTGCGCGGCAGGCGCGAGCCTCCAGTGGCAGATAGCA 
                 
                 
                   CCACATCCCAGGCCTTCCCTCTGAGGGCTGGGGGAAATGGGCAGCTCCAGTATTGGC 
                 
                 
                   CATTTTCTAGTTCAGACCTGTACAACTGGAAGAACAATAACCCCTCTTTCAGTGAGGA 
                 
                 
                   CCCCGGCAAACTGACCGCCCTGATCGAATCCGTGCTGATTACCCATCAGCCCACATG 
                 
                 
                   GGACGATTGTCAGCAGCTCCTGGGCACCCTGCTGACCGGAGAGGAAAAGCAGCGCGT 
                 
                 
                   GCTGCTGGAGGCTCGCAAAGCAGTCCGAGGGGACGATGGACGGCCCACACAGCTCCC 
                 
                 
                   TAATGAGGTGGACGCCGCTTTTCCACTGGAAAGACCCGACTGGGATTATACTACCCA 
                 
                 
                   GGCAGGGAGAAACCACCTGGTCCATTACAGGCAGCTCCTGCTGGCAGGCCTGCAGAA 
                 
                 
                   TGCCGGGAGATCCCCCACCAACCTGGCCAAGGTGAAAGGCATCACACAGGGGCCTAA 
                 
                 
                   TGAGTCACCAAGCGCCTTTCTGGAGAGGCTGAAGGAAGCTTACCGACGGTATACCCC 
                 
                 
                   ATACGACCCTGAGGACCCCGGACAGGAAACAAACGTCTCCATGTCTTTCATCTGGCA 
                 
                 
                   GTCTGCCCCAGACATTGGGCGGAAGCTGGAGAGACTGGAAGACCTGAAGAACAAGA 
                 
                 
                   CCCTGGGCGACCTGGTGCGGGAGGCTGAAAAGATCTTCAACAAACGGGAGACCCCCG 
                 
                 
                   AGGAAAGAGAGGAAAGGATTAGAAGGGAAACTGAGGAAAAGGAGGAACGCCGACG 
                 
                 
                   GACCGAGGACGAACAGAAGGAGAAAGAACGAGATCGGCGGCGGCACCGGGAGATG 
                 
                 
                   TCAAAGCTGCTGGCCACCGTGGTCAGCGGACAGAAACAGGACAGACAGGGAGGAGA 
                 
                 
                   GCGACGGAGAAGCCAGCTCGACAGGGATCAGTGCGCATACTGTAAGGAAAAAGGCC 
                 
                 
                   ATTGGGCCAAGGATTGCCCCAAAAAGCCAAGAGGACCAAGAGGACCAAGACCACAG 
                 
                 
                   ACATCACTGCTGACCCTGGACGAC 
                 
                 
                     
                 
                 
                   SEQ ID NO: 4 is a SARS-CoV-2 Spike Glycoprotein, Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLAL 
                 
                 
                   HRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTL 
                 
                 
                   KSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVAD 
                 
                 
                   YSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYK 
                 
                 
                   LPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVE 
                 
                 
                   GFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNF 
                 
                 
                   NGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTS 
                 
                 
                   NQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDI 
                 
                 
                   PIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILP 
                 
                 
                   VSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQI 
                 
                 
                   YKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC 
                 
                 
                   AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGV 
                 
                 
                   TQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFG 
                 
                 
                   AISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSEC 
                 
                 
                   VLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPR 
                 
                 
                   EGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEEL 
                 
                 
                   DKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWP 
                 
                 
                   WYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT 
                 
                 
                     
                 
                 
                   SEQ ID NO: 5 is a SARS-CoV-2 Spike Glycoprotein, Nucleotide Sequence 
                 
                 
                   (Wuhan-Hu-1: Genbank Ref: MN908947) 
                 
                 
                   ATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAAC 
                 
                 
                   CAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCT 
                 
                 
                   GACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTT 
                 
                 
                   TTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGG 
                 
                 
                   TTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTC 
                 
                 
                   TAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTA 
                 
                 
                   CTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAA 
                 
                 
                   TGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGA 
                 
                 
                   GTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTC 
                 
                 
                   TTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTA 
                 
                 
                   AGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCG 
                 
                 
                   TGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATT 
                 
                 
                   AACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTG 
                 
                 
                   ATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCT 
                 
                 
                   AGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGT 
                 
                 
                   GCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAA 
                 
                 
                   GGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTC 
                 
                 
                   CTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCT 
                 
                 
                   GTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTAT 
                 
                 
                   ATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAAT 
                 
                 
                   GATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCA 
                 
                 
                   GACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAG 
                 
                 
                   ATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGG 
                 
                 
                   TGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAG 
                 
                 
                   AGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAA 
                 
                 
                   GGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGG 
                 
                 
                   TTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACT 
                 
                 
                   GTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACT 
                 
                 
                   TCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTT 
                 
                 
                   CCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGAC 
                 
                 
                   ACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCA 
                 
                 
                   GGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAG 
                 
                 
                   TCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGT 
                 
                 
                   TCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACT 
                 
                 
                   CATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGAC 
                 
                 
                   TAATTCTCCTCGGCGGGCACGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATG 
                 
                 
                   TCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAA 
                 
                 
                   ATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGT 
                 
                 
                   AGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAA 
                 
                 
                   TATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAG 
                 
                 
                   ACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAA 
                 
                 
                   TTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAG 
                 
                 
                   CAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGC 
                 
                 
                   TTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTG 
                 
                 
                   CACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGC 
                 
                 
                   TCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCA 
                 
                 
                   GGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTG 
                 
                 
                   GAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATA 
                 
                 
                   GTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACT 
                 
                 
                   TCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAG 
                 
                 
                   CTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACAAAG 
                 
                 
                   TTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGA 
                 
                 
                   CATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGC 
                 
                 
                   TGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGA 
                 
                 
                   AAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCA 
                 
                 
                   TGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCAT 
                 
                 
                   GATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGT 
                 
                 
                   TTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGT 
                 
                 
                   GTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTG 
                 
                 
                   CAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACA 
                 
                 
                   TCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTC 
                 
                 
                   AAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCG 
                 
                 
                   ATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATGGCCATGGTACATTTGGC 
                 
                 
                   TAGGTTTTATAGCTGGCTTGATTGCCATAGTAATGGTGACAATTATGCTTTGCTGTAT 
                 
                 
                   GACCAGTTGCTGTAGTTGTCTCAAGGGCTGTTGTTCTTGTGGATCCTGCTGCAAATTT 
                 
                 
                   GATGAAGACGACTCTGAGCCAGTGCTCAAAGGAGTCAAATTACATTACACATAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 6 is a SARS-CoV-2 Spike Glycoprotein, Nucleotide Sequence, 
                 
                 
                   Codon Optimized For Expression In Human Cell 
                 
                 
                   ATGTTCGTGTTTCTGGTGCTGCTGCCTCTGGTGAGCTCCCAGTGCGTGAACCTGACCA 
                 
                 
                   CAAGGACCCAGCTCCCCCCTGCCTATACCAATTCCTTCACACGGGGCGTGTACTATCC 
                 
                 
                   CGACAAGGTGTTTAGATCTAGCGTGCTGCACTCCACACAGGATCTGTTTCTGCCTTTC 
                 
                 
                   TTTTCTAACGTGACCTGGTTCCACGCCATCCATGTGAGCGGCACCAATGGCACAAAGC 
                 
                 
                   GGTTCGACAATCCAGTGCTGCCCTTTAACGATGGCGTGTACTTCGCCTCCACCGAGAA 
                 
                 
                   GTCTAACATCATCAGAGGCTGGATCTTTGGCACCACACTGGACAGCAAGACACAGTC 
                 
                 
                   CCTGCTGATCGTGAACAATGCCACCAACGTGGTCATCAAGGTGTGCGAGTTCCAGTTT 
                 
                 
                   TGTAATGATCCATTCCTGGGCGTGTACTATCACAAGAACAATAAGTCTTGGATGGAG 
                 
                 
                   AGCGAGTTTCGCGTGTATTCCTCTGCCAACAATTGCACATTTGAGTACGTGTCCCAGC 
                 
                 
                   CCTTCCTGATGGACCTGGAGGGCAAGCAGGGCAATTTCAAGAACCTGAGGGAGTTCG 
                 
                 
                   TGTTTAAGAATATCGATGGCTACTTCAAGATCTACTCCAAGCACACCCCAATCAACCT 
                 
                 
                   GGTGCGCGACCTGCCACAGGGCTTCTCTGCCCTGGAGCCACTGGTGGATCTGCCCATC 
                 
                 
                   GGCATCAACATCACCCGGTTTCAGACACTGCTGGCCCTGCACAGAAGCTACCTGACA 
                 
                 
                   CCAGGCGACAGCTCCTCTGGATGGACCGCCGGGGCCGCCGCCTACTATGTGGGCTAT 
                 
                 
                   CTGCAGCCCAGGACCTTCCTGCTGAAGTACAACGAGAATGGCACCATCACAGACGCA 
                 
                 
                   GTGGATTGCGCCCTGGACCCCCTGTCTGAGACCAAGTGTACACTGAAGAGCTTTACC 
                 
                 
                   GTGGAGAAGGGCATCTATCAGACAAGCAATTTCAGGGTGCAGCCTACCGAGTCCATC 
                 
                 
                   GTGCGCTTTCCCAATATCACAAACCTGTGCCCTTTTGGCGAGGTGTTCAACGCAACCA 
                 
                 
                   GGTTCGCCAGCGTGTACGCATGGAATAGGAAGCGCATCTCCAACTGCGTGGCCGACT 
                 
                 
                   ATTCTGTGCTGTACAACAGCGCCTCCTTCTCTACCTTTAAGTGCTATGGCGTGAGCCC 
                 
                 
                   CACAAAGCTGAATGACCTGTGCTTTACCAACGTGTACGCCGATTCCTTCGTGATCAGG 
                 
                 
                   GGCGACGAGGTGCGCCAGATCGCACCAGGACAGACAGGCAAGATCGCAGACTACAA 
                 
                 
                   TTATAAGCTGCCTGACGATTTCACCGGCTGCGTGATCGCCTGGAACTCTAACAATCTG 
                 
                 
                   GATAGCAAAGTGGGCGGCAACTACAATTATCTGTACCGGCTGTTTAGAAAGTCTAAT 
                 
                 
                   CTGAAGCCATTCGAGAGGGACATCTCCACAGAGATCTACCAGGCCGGCTCTACCCCC 
                 
                 
                   TGCAATGGCGTGGAGGGCTTTAACTGTTATTTCCCTCTGCAGAGCTACGGCTTCCAGC 
                 
                 
                   CAACAAACGGCGTGGGCTATCAGCCCTACCGCGTGGTGGTGCTGTCTTTTGAGCTGCT 
                 
                 
                   GCACGCACCTGCAACAGTGTGCGGACCAAAGAAGAGCACCAATCTGGTGAAGAACA 
                 
                 
                   AGTGCGTGAACTTCAACTTCAACGGCCTGACCGGCACAGGCGTGCTGACCGAGTCCA 
                 
                 
                   ACAAGAAGTTCCTGCCTTTTCAGCAGTTCGGCAGGGACATCGCAGATACCACAGACG 
                 
                 
                   CCGTGCGCGACCCTCAGACCCTGGAGATCCTGGACATCACACCATGCTCCTTCGGCG 
                 
                 
                   GCGTGTCTGTGATCACACCAGGCACCAATACAAGCAACCAGGTGGCCGTGCTGTATC 
                 
                 
                   AGGACGTGAATTGTACCGAGGTGCCCGTGGCCATCCACGCAGATCAGCTCACCCCTA 
                 
                 
                   CATGGCGGGTGTACTCTACCGGCAGCAACGTGTTCCAGACAAGAGCCGGCTGCCTGA 
                 
                 
                   TCGGAGCCGAGCATGTGAACAATAGCTATGAGTGCGACATCCCTATCGGAGCCGGCA 
                 
                 
                   TCTGTGCCTCCTACCAGACCCAGACAAACTCCCCACGGAGAGCCCGGTCTGTGGCCA 
                 
                 
                   GCCAGTCCATCATCGCCTATACCATGAGCCTGGGGGCCGAGAACAGCGTGGCCTACT 
                 
                 
                   CCAACAATTCTATCGCCATCCCTACCAACTTCACAATCTCCGTGACCACAGAGATCCT 
                 
                 
                   GCCAGTGAGCATGACCAAGACATCCGTGGACTGCACAATGTATATCTGTGGCGATTC 
                 
                 
                   CACCGAGTGCTCTAACCTGCTGCTGCAGTACGGCTCTTTTTGTACCCAGCTCAACAGA 
                 
                 
                   GCCCTGACAGGCATCGCCGTGGAGCAGGACAAGAACACACAGGAGGTGTTCGCCCA 
                 
                 
                   GGTGAAGCAGATCTACAAGACCCCACCCATCAAGGACTTTGGCGGCTTCAACTTCAG 
                 
                 
                   CCAGATCCTGCCCGATCCTAGCAAGCCATCCAAGCGGTCTTTTATCGAGGACCTGCTG 
                 
                 
                   TTCAACAAGGTGACCCTGGCCGATGCCGGCTTCATCAAGCAGTATGGCGATTGCCTG 
                 
                 
                   GGCGACATCGCCGCCAGAGACCTGATCTGTGCCCAGAAGTTTAATGGCCTGACCGTG 
                 
                 
                   CTGCCTCCACTGCTGACAGATGAGATGATCGCCCAGTACACATCTGCCCTGCTGGCCG 
                 
                 
                   GCACCATCACAAGCGGATGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCCTTTG 
                 
                 
                   CCATGCAGATGGCCTATCGGTTCAACGGCATCGGCGTGACCCAGAATGTGCTGTACG 
                 
                 
                   AGAACCAGAAGCTGATCGCCAATCAGTTTAACTCCGCCATCGGCAAGATCCAGGACT 
                 
                 
                   CTCTGAGCTCCACAGCCAGCGCCCTGGGCAAGCTGCAGGATGTGGTGAATCAGAACG 
                 
                 
                   CCCAGGCCCTGAATACCCTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAGCA 
                 
                 
                   GCGTGCTGAACGACATCCTGAGCCGGCTGGACAAGGTGGAGGCAGAGGTGCAGATC 
                 
                 
                   GACCGGCTGATCACAGGCAGACTGCAGTCCCTGCAGACCTACGTGACACAGCAGCTC 
                 
                 
                   ATCAGGGCCGCCGAGATCAGGGCCTCTGCCAATCTGGCCGCCACCAAGATGAGCGAG 
                 
                 
                   TGCGTGCTGGGCCAGTCCAAGAGAGTGGACTTTTGTGGCAAGGGCTATCACCTGATG 
                 
                 
                   AGCTTCCCACAGTCCGCCCCTCACGGAGTGGTGTTTCTGCATGTGACCTACGTGCCAG 
                 
                 
                   CCCAGGAGAAGAACTTCACCACAGCCCCCGCAATCTGCCACGATGGCAAGGCACACT 
                 
                 
                   TTCCCCGGGAGGGCGTGTTCGTGAGCAACGGCACCCACTGGTTTGTGACACAGCGCA 
                 
                 
                   ATTTCTACGAGCCACAGATCATCACCACAGACAATACATTCGTGTCCGGCAACTGTG 
                 
                 
                   ACGTGGTCATCGGCATCGTGAACAATACCGTGTATGATCCTCTGCAGCCAGAGCTGG 
                 
                 
                   ACTCTTTTAAGGAGGAGCTGGATAAGTACTTCAAGAATCACACCAGCCCCGACGTGG 
                 
                 
                   ATCTGGGCGACATCTCTGGCATCAATGCCAGCGTGGTGAACATCCAGAAGGAGATCG 
                 
                 
                   ACAGGCTGAACGAGGTGGCCAAGAATCTGAACGAGTCCCTGATCGATCTGCAGGAGC 
                 
                 
                   TGGGCAAGTATGAGCAGTACATCAAGTGGCCCTGGTATATCTGGCTGGGCTTCATCG 
                 
                 
                   CCGGCCTGATCGCCATCGTGATGGTGACCATCATGCTGTGCTGTATGACAAGCTGCTG 
                 
                 
                   TTCCTGCCTGAAGGGCTGCTGTTCTTGTGGCTCCTGCTGTAAGTTTGATGAGGACGAT 
                 
                 
                   AGCGAGCCTGTGCTGAAGGGCGTGAAGCTGCACTACACCTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 7 is a SARS-CoV Spike Glycoprotein, Amino Acid Sequence 
                 
                 
                   (HKU-39849, Genbank Ref: JN854286.1) 
                 
                 
                   MFIFLLFLTLTSGSDLDRCTTFDDVQAPNYTQHTSSMRGVYYPDEIFRSDTLYLTQDLFLP 
                 
                 
                   FYSNVTGFHTINHTFGNPVIPFKDGIYFAATEKSNVVRGWVFGSTMNNKSQSVIIINNSTN 
                 
                 
                   VVIRACNFELCDNPFFAVSKPMGTQTHTMIFDNAFNCTFEYISDAFSLDVSEKSGNFKHLR 
                 
                 
                   EFVFKNKDGFLYVYKGYQPIDVVRDLPSGFNTLKPIFKLPLGINITNFRAILTAFSPAQDIW 
                 
                 
                   GTSAAAYFVGYLKPTTFMLKYDENGTITDAVDCSQNPLAELKCSVKSFEIDKGIYQTSNF 
                 
                 
                   RVVPSGDVVRFPNITNLCPFGEVFNATKFPSVYAWERKKISNCVADYSVLYNSTFFSTFKC 
                 
                 
                   YGVSATKLNDLCFSNVYADSFVVKGDDVRQIAPGQTGVIADYNYKLPDDFMGCVLAWN 
                 
                 
                   TRNIDATSTGNYNYKYRYLRHGKLRPFERDISNVPFSPDGKPCTPPALNCYWPLNDYGFY 
                 
                 
                   TTTGIGYQPYRVVVLSFELLNAPATVCGPKLSTDLIKNQCVNFNFNGLTGTGVLTPSSKRF 
                 
                 
                   QPFQQFGRDVSDFTDSVRDPKTSEILDISPCSFGGVSVITPGTNASSEVAVLYQDVNCTDV 
                 
                 
                   STAIHADQLTPAWRIYSTGNNVFQTQAGCLIGAEHVDTSYECDIPIGAGICASYHTVSLLR 
                 
                 
                   STSQKSIVAYTMSLGADSSIAYSNNTIAIPTNFSISITTEVMPVSMAKTSVDCNMYICGDST 
                 
                 
                   ECANLLLQYGSFCTQLNRALSGIAAEQDRNTREVFAQVKQMYKTPTLKYFGGFNFSQILP 
                 
                 
                   DPLKPTKRSFIEDLLFNKVTLADAGFMKQYGECLGDINARDLICAQKFNGLTVLPPLLTD 
                 
                 
                   DMIAAYTAALVSGTATAGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKQIAN 
                 
                 
                   QFNKAISQIQESLTTTSTALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDK 
                 
                 
                   VEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKG 
                 
                 
                   YHLMSFPQAAPHGVVFLHVTYVPSQERNFTTAPAICHEGKAYFPREGVFVFNGTSWFITQ 
                 
                 
                   RNFFSPQIITTDNTFVSGNCDVVIGIINNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGD 
                 
                 
                   ISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYVWLGFIAGLIAIVM 
                 
                 
                   VTILLCCMTSCCSCLKGACSCGSCCKFDEDDSEPVLKGVKLHYT 
                 
                 
                     
                 
                 
                   SEQ ID NO: 8 is a SARS-CoV Spike Glycoprotein, Nucleotide Sequence 
                 
                 
                   ATGTTTATTTTCTTATTATTTCTTACTCTCACTAGTGGTAGTGACCTTGACCGGTGCAC 
                 
                 
                   CACTTTTGATGATGTTCAAGCTCCTAATTACACTCAACATACTTCATCTATGAGGGGG 
                 
                 
                   GTTTACTATCCTGATGAAATTTTTAGATCAGACACTCTTTATTTAACTCAGGATTTATT 
                 
                 
                   TCTTCCATTTTATTCTAATGTTACAGGGTTTCATACTATTAATCATACGTTTGGCAACC 
                 
                 
                   CTGTCATACCTTTTAAGGATGGTATTTATTTTGCTGCCACAGAGAAATCAAATGTTGT 
                 
                 
                   CCGTGGTTGGGTTTTTGGTTCTACCATGAACAACAAGTCACAGTCGGTGATTATTATT 
                 
                 
                   AACAATTCTACTAATGTTGTTATACGAGCATGTAACTTTGAATTGTGTGACAACCCTT 
                 
                 
                   TCTTTGCTGTTTCTAAACCCATGGGTACACAGACACATACTATGATATTCGATAATGC 
                 
                 
                   ATTTAATTGCACTTTCGAGTACATATCTGATGCCTTTTCGCTTGATGTTTCAGAAAAGT 
                 
                 
                   CAGGTAATTTTAAACACTTACGAGAGTTTGTGTTTAAAAATAAAGATGGGTTTCTCTA 
                 
                 
                   TGTTTATAAGGGCTATCAACCTATAGATGTAGTTCGTGATCTACCTTCTGGTTTTAAC 
                 
                 
                   ACTTTGAAACCTATTTTTAAGTTGCCTCTTGGTATTAACATTACAAATTTTAGAGCCAT 
                 
                 
                   TCTTACAGCCTTTTCACCTGCTCAAGACATTTGGGGCACGTCAGCTGCAGCCTATTTT 
                 
                 
                   GTTGGCTATTTAAAGCCAACTACATTTATGCTCAAGTATGATGAAAATGGTACAATCA 
                 
                 
                   CAGATGCTGTTGATTGTTCTCAAAATCCACTTGCTGAACTCAAATGCTCTGTTAAGAG 
                 
                 
                   CTTTGAGATTGACAAAGGAATTTACCAGACCTCTAATTTCAGGGTTGTTCCCTCAGGA 
                 
                 
                   GATGTTGTGAGATTCCCTAATATTACAAACTTGTGTCCTTTTGGAGAGGTTTTTAATG 
                 
                 
                   CTACTAAATTCCCTTCTGTCTATGCATGGGAGAGAAAAAAAATTTCTAATTGTGTTGC 
                 
                 
                   TGATTACTCTGTGCTCTACAACTCAACATTTTTTTCAACCTTTAAGTGCTATGGCGTTT 
                 
                 
                   CTGCCACTAAGTTGAATGATCTTTGCTTCTCCAATGTCTATGCAGATTCTTTTGTAGTC 
                 
                 
                   AAGGGAGATGATGTAAGACAAATAGCGCCAGGACAAACTGGTGTTATTGCTGATTAT 
                 
                 
                   AATTATAAATTGCCAGATGATTTCATGGGTTGTGTCCTTGCTTGGAATACTAGGAACA 
                 
                 
                   TTGATGCTACTTCAACTGGTAATTATAATTATAAATATAGGTATCTTAGACATGGCAA 
                 
                 
                   GCTTAGGCCCTTTGAGAGAGACATATCTAATGTGCCTTTCTCCCCTGATGGCAAACCT 
                 
                 
                   TGCACCCCACCTGCTCTTAATTGTTATTGGCCATTAAATGATTATGGTTTTTACACCAC 
                 
                 
                   TACTGGCATTGGCTACCAACCTTACAGAGTTGTAGTACTTTCTTTTGAACTTTTAAATG 
                 
                 
                   CACCGGCCACGGTTTGTGGACCAAAATTATCCACTGACCTTATTAAGAACCAGTGTGT 
                 
                 
                   CAATTTTAATTTTAATGGACTCACTGGTACTGGTGTGTTAACTCCTTCTTCAAAGAGA 
                 
                 
                   TTTCAACCATTTCAACAATTTGGCCGTGATGTTTCTGATTTCACTGATTCCGTTCGAGA 
                 
                 
                   TCCTAAAACATCTGAAATATTAGACATTTCACCTTGCTCTTTTGGGGGTGTAAGTGTA 
                 
                 
                   ATTACACCTGGAACAAATGCTTCATCTGAAGTTGCTGTTCTATATCAAGATGTTAACT 
                 
                 
                   GCACTGATGTTTCTACAGCAATTCATGCAGATCAACTCACACCAGCTTGGCGCATATA 
                 
                 
                   TTCTACTGGAAACAATGTATTCCAGACTCAAGCAGGCTGTCTTATAGGAGCTGAGCAT 
                 
                 
                   GTCGACACTTCTTATGAGTGCGACATTCCTATTGGAGCTGGCATTTGTGCTAGTTACC 
                 
                 
                   ATACAGTTTCTTTATTACGTAGTACTAGCCAAAAATCTATTGTGGCTTATACTATGTCT 
                 
                 
                   TTAGGTGCTGATAGTTCAATTGCTTACTCTAATAACACCATTGCTATACCTACTAACTT 
                 
                 
                   TTCAATTAGCATTACTACAGAAGTAATGCCTGTTTCTATGGCTAAAACCTCCGTAGAT 
                 
                 
                   TGTAATATGTACATCTGCGGAGATTCTACTGAATGTGCTAATTTGCTTCTCCAATATG 
                 
                 
                   GTAGCTTTTGCACACAACTAAATCGTGCACTCTCAGGTATTGCTGCTGAACAGGATCG 
                 
                 
                   CAACACACGTGAAGTGTTCGCTCAAGTCAAACAAATGTACAAAACCCCAACTTTGAA 
                 
                 
                   ATATTTTGGTGGTTTTAATTTTTCACAAATATTACCTGACCCTCTAAAGCCAACTAAG 
                 
                 
                   AGGTCTTTTATTGAGGACTTGCTCTTTAATAAGGTGACACTCGCTGATGCTGGCTTCA 
                 
                 
                   TGAAGCAATATGGCGAATGCCTAGGTGATATTAATGCTAGAGATCTCATTTGTGCGC 
                 
                 
                   AGAAGTTCAATGGACTTACAGTGTTGCCACCTCTGCTCACTGATGATATGATTGCTGC 
                 
                 
                   CTACACTGCTGCTCTAGTTAGTGGTACTGCCACTGCTGGATGGACATTTGGTGCTGGC 
                 
                 
                   GCTGCTCTTCAAATACCTTTTGCTATGCAAATGGCATATAGGTTCAATGGCATTGGAG 
                 
                 
                   TTACCCAAAATGTTCTCTATGAGAACCAAAAACAAATCGCCAACCAATTTAACAAGG 
                 
                 
                   CGATTAGTCAAATTCAAGAATCACTTACAACAACATCAACTGCATTGGGCAAGCTGC 
                 
                 
                   AAGACGTTGTTAACCAGAATGCTCAAGCATTAAACACACTTGTTAAACAACTTAGCT 
                 
                 
                   CTAATTTTGGTGCAATTTCAAGTGTGCTAAATGATATCCTTTCGCGACTTGATAAAGT 
                 
                 
                   CGAGGCGGAGGTACAAATTGACAGGTTAATTACAGGCAGACTTCAAAGCCTTCAAAC 
                 
                 
                   CTATGTAACACAACAACTAATCAGGGCTGCTGAAATCAGGGCTTCTGCTAATCTTGCT 
                 
                 
                   GCTACTAAAATGTCTGAGTGTGTTCTTGGACAATCAAAAAGAGTTGACTTTTGTGGAA 
                 
                 
                   AGGGCTACCACCTTATGTCCTTCCCACAAGCAGCCCCGCATGGTGTTGTCTTCCTACA 
                 
                 
                   TGTCACGTATGTGCCATCCCAGGAGAGGAACTTCACCACAGCGCCAGCAATTTGTCA 
                 
                 
                   TGAAGGCAAAGCATACTTCCCTCGTGAAGGTGTTTTTGTGTTTAATGGCACTTCTTGG 
                 
                 
                   TTTATTACACAGAGGAACTTCTTTTCTCCACAAATAATTACTACAGACAATACATTTG 
                 
                 
                   TCTCAGGAAATTGTGATGTCGTTATTGGCATCATTAACAACACAGTTTATGATCCTCT 
                 
                 
                   GCAACCTGAGCTTGACTCATTCAAAGAAGAGCTGGACAAGTACTTCAAAAATCATAC 
                 
                 
                   ATCACCAGATGTTGATCTTGGCGACATTTCAGGCATTAACGCTTCTGTCGTCAACATT 
                 
                 
                   CAAAAAGAAATTGACCGCCTCAATGAGGTCGCTAAAAATTTAAATGAATCACTCATT 
                 
                 
                   GACCTTCAAGAATTGGGAAAATATGAGCAATATATTAAATGGCCTTGGTATGTTTGG 
                 
                 
                   CTCGGCTTCATTGCTGGACTAATTGCCATCGTCATGGTTACAATCTTGCTTTGTTGCAT 
                 
                 
                   GACTAGTTGTTGCAGTTGCCTCAAGGGTGCATGCTCTTGTGGTTCTTGCTGCAAGTTT 
                 
                 
                   GATGAGGATGACTCTGAGCCAGTTCTCAAGGGTGTCAAATTACATTACACATAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 9 is a SARS-CoV Spike Glycoprotein, Nucleotide Sequence, 
                 
                 
                   Codon Optimized for Expression In Human Cells 
                 
                 
                   ATGTTCATCTTTCTGCTGTTCCTGACCCTGACAAGCGGCTCCGACCTGGATAGGTGCA 
                 
                 
                   CCACATTTGACGATGTGCAGGCCCCCAACTACACACAGCACACCAGCTCCATGAGGG 
                 
                 
                   GCGTGTACTATCCTGATGAGATCTTCCGCTCTGACACACTGTACCTGACCCAGGACCT 
                 
                 
                   GTTCCTGCCTTTTTATAGCAATGTGACAGGCTTCCACACCATCAATCACACATTTGGC 
                 
                 
                   AACCCCGTGATCCCTTTCAAGGATGGCATCTACTTTGCCGCCACCGAGAAGTCTAACG 
                 
                 
                   TGGTGCGGGGATGGGTGTTCGGCAGCACAATGAACAATAAGTCTCAGAGCGTGATCA 
                 
                 
                   TCATCAACAATAGCACCAACGTGGTCATCAGAGCCTGCAATTTTGAGCTGTGCGACA 
                 
                 
                   ACCCCTTCTTTGCCGTGTCCAAGCCTATGGGCACCCAGACACACACCATGATCTTTGA 
                 
                 
                   TAATGCCTTCAACTGTACCTTTGAGTACATCAGCGATGCCTTTTCCCTGGACGTGTCT 
                 
                 
                   GAGAAGTCCGGCAACTTCAAGCACCTGAGGGAGTTCGTGTTTAAGAATAAGGACGGC 
                 
                 
                   TTCCTGTACGTGTATAAGGGCTATCAGCCCATCGATGTGGTGCGCGACCTGCCTTCCG 
                 
                 
                   GCTTCAACACCCTGAAGCCAATCTTTAAGCTGCCCCTGGGCATCAATATCACCAACTT 
                 
                 
                   CAGGGCCATCCTGACAGCCTTTAGCCCAGCACAGGACATCTGGGGCACCAGCGCCGC 
                 
                 
                   CGCCTACTTCGTGGGCTATCTGAAGCCCACCACCTTCATGCTGAAGTACGATGAGAAC 
                 
                 
                   GGCACAATCACCGACGCCGTGGATTGCAGCCAGAATCCACTGGCCGAGCTGAAGTGT 
                 
                 
                   TCCGTGAAGTCTTTCGAGATCGACAAGGGCATCTATCAGACCTCCAACTTTAGGGTGG 
                 
                 
                   TGCCATCTGGCGATGTGGTGCGCTTCCCAAATATCACCAACCTGTGCCCCTTCGGCGA 
                 
                 
                   GGTGTTTAATGCCACAAAGTTCCCCAGCGTGTACGCCTGGGAGCGCAAGAAGATCAG 
                 
                 
                   CAACTGCGTGGCCGACTACTCCGTGCTGTATAATAGCACCTTCTTCAGCACCTTCAAG 
                 
                 
                   TGCTACGGCGTGAGCGCCACCAAGCTGAATGACCTGTGCTTCTCTAACGTGTATGCCG 
                 
                 
                   ATAGCTTTGTGGTGAAGGGCGACGATGTGAGGCAGATCGCACCTGGACAGACCGGCG 
                 
                 
                   TGATCGCAGACTACAACTATAAGCTGCCAGACGATTTCATGGGCTGCGTGCTGGCCT 
                 
                 
                   GGAATACACGCAACATCGATGCCACATCCACCGGCAACTACAATTATAAGTACCGGT 
                 
                 
                   ATCTGAGACACGGCAAGCTGCGGCCCTTCGAGAGAGACATCTCCAATGTGCCATTTT 
                 
                 
                   CTCCAGATGGCAAGCCATGCACCCCACCTGCCCTGAATTGTTACTGGCCTCTGAACGA 
                 
                 
                   CTACGGCTTCTATACCACAACCGGCATCGGCTACCAGCCTTATAGGGTGGTGGTGCTG 
                 
                 
                   TCCTTTGAGCTGCTGAACGCACCTGCAACCGTGTGCGGACCAAAGCTGTCTACAGATC 
                 
                 
                   TGATCAAGAATCAGTGCGTGAACTTCAACTTCAACGGCCTGACAGGCACCGGCGTGC 
                 
                 
                   TGACCCCTTCTAGCAAGCGGTTCCAGCCATTTCAGCAGTTCGGCAGAGACGTGAGCG 
                 
                 
                   ATTTCACCGACTCCGTGCGCGACCCAAAGACATCCGAGATCCTGGACATCAGCCCCT 
                 
                 
                   GCTCCTTTGGCGGCGTGTCTGTGATCACACCTGGCACCAACGCCTCCTCTGAGGTGGC 
                 
                 
                   CGTGCTGTACCAGGATGTGAATTGTACCGACGTGAGCACAGCAATCCACGCAGACCA 
                 
                 
                   GCTCACCCCAGCATGGCGGATCTATTCCACCGGCAACAACGTGTTCCAGACACAGGC 
                 
                 
                   AGGATGCCTGATCGGAGCCGAGCATGTGGATACAAGCTACGAGTGCGACATCCCCAT 
                 
                 
                   CGGAGCCGGCATCTGTGCCTCTTATCACACCGTGAGCCTGCTGAGATCCACATCTCAG 
                 
                 
                   AAGTCTATCGTGGCCTACACCATGAGCCTGGGGGCCGATAGCTCCATCGCCTATTCCA 
                 
                 
                   ACAATACCATCGCCATCCCAACAAACTTCAGCATCTCCATCACAACCGAAGTGATGC 
                 
                 
                   CCGTGTCCATGGCCAAGACCTCTGTGGACTGCAACATGTACATCTGTGGCGATAGCA 
                 
                 
                   CAGAGTGCGCCAATCTGCTGCTGCAGTATGGCTCCTTTTGTACCCAGCTCAACCGGGC 
                 
                 
                   CCTGTCTGGAATCGCCGCCGAGCAGGACAGGAATACACGCGAGGTGTTCGCCCAGGT 
                 
                 
                   GAAGCAGATGTACAAGACACCTACCCTGAAGTATTTTGGCGGCTTCAACTTTTCTCAG 
                 
                 
                   ATCCTGCCTGATCCACTGAAGCCAACCAAGCGGAGCTTCATCGAGGACCTGCTGTTTA 
                 
                 
                   ATAAGGTGACACTGGCCGATGCCGGCTTCATGAAGCAGTACGGCGAGTGCCTGGGCG 
                 
                 
                   ACATCAACGCCAGAGACCTGATCTGTGCCCAGAAGTTTAATGGCCTGACCGTGCTGC 
                 
                 
                   CACCCCTGCTGACAGACGATATGATCGCAGCATATACCGCCGCCCTGGTGTCCGGCA 
                 
                 
                   CAGCCACCGCCGGCTGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCTTTCGCCA 
                 
                 
                   TGCAGATGGCCTACCGGTTTAACGGCATCGGCGTGACCCAGAATGTGCTGTATGAGA 
                 
                 
                   ACCAGAAGCAGATCGCCAATCAGTTTAACAAGGCCATCAGCCAGATCCAGGAGTCCC 
                 
                 
                   TGACAACCACATCTACCGCCCTGGGCAAGCTGCAGGACGTGGTGAATCAGAACGCCC 
                 
                 
                   AGGCCCTGAATACACTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAGCAGCG 
                 
                 
                   TGCTGAACGACATCCTGAGCCGGCTGGACAAGGTGGAGGCAGAGGTGCAGATCGAT 
                 
                 
                   AGGCTGATCACCGGCAGACTGCAGTCTCTGCAGACATACGTGACCCAGCAGCTCATC 
                 
                 
                   AGGGCCGCCGAGATCAGAGCCAGCGCCAACCTGGCCGCCACAAAGATGTCCGAGTG 
                 
                 
                   CGTGCTGGGCCAGTCTAAGAGGGTGGACTTCTGTGGCAAGGGCTACCACCTGATGTC 
                 
                 
                   CTTTCCACAGGCCGCCCCTCACGGAGTGGTGTTCCTGCATGTGACCTATGTGCCTTCT 
                 
                 
                   CAGGAGCGCAACTTTACCACAGCCCCAGCAATCTGCCACGAGGGCAAGGCATACTTC 
                 
                 
                   CCCCGGGAGGGCGTGTTCGTGTTTAACGGCACCTCCTGGTTTATCACACAGAGAAATT 
                 
                 
                   TCTTTTCCCCTCAGATCATCACCACAGACAATACCTTCGTGAGCGGCAACTGTGACGT 
                 
                 
                   GGTCATCGGCATCATCAACAATACAGTGTACGATCCTCTGCAGCCAGAGCTGGACAG 
                 
                 
                   CTTCAAGGAGGAGCTGGATAAGTACTTCAAGAACCACACCTCCCCCGACGTGGATCT 
                 
                 
                   GGGCGACATCAGCGGCATCAATGCCTCCGTGGTGAACATCCAGAAGGAGATCGACAG 
                 
                 
                   ACTGAATGAGGTGGCCAAGAATCTGAACGAGTCCCTGATCGATCTGCAGGAGCTGGG 
                 
                 
                   CAAGTACGAGCAGTATATCAAGTGGCCATGGTACGTGTGGCTGGGCTTCATCGCCGG 
                 
                 
                   CCTGATCGCCATCGTGATGGTGACCATCCTGCTGTGCTGTATGACATCTTGCTGTAGC 
                 
                 
                   TGCCTGAAGGGAGCCTGCTCCTGTGGCTCTTGCTGTAAGTTTGACGAGGACGATAGC 
                 
                 
                   GAGCCCGTGCTGAAGGGCGTGAAGCTGCACTATACCTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 10 is a MERS-CoV Spike Glycoprotein, Amino Acid Sequence 
                 
                 
                   MIHSVFLLMFLLTPTESYVDVGPDSVKSACIEVDIQQTFFDKTWPRPIDVSKADGITYPQGR 
                 
                 
                   TYSNITITYQGLFPYQGDHGDMYVYSAGHATGTTPQKLFVANYSQDVKQFANGFVVRIG 
                 
                 
                   AAANSTGTVIISPSTSATIRKIYPAFMLGSSVGNFSDGKMGRFFNHTLVLLPDGCGTLLRA 
                 
                 
                   FYCILEPRSGNHCPAGNSYTSFATYHTPATDCSDGNYNRNASLNSFKEYFNLRNCTFMYT 
                 
                 
                   YNITEDEILEWFGITQTAQGVHLFSSRYVDLYGGNMFQFATLPVYDTIKYYSIIPHSIRSIQS 
                 
                 
                   DRKAWAAFYVYKLQPLTFLLDFSVDGYIRRAIDCGFNDLSQLHCSYESFDVESGVYSVSS 
                 
                 
                   FEAKPSGSVVEQAEGVECDFSPLLSGTPPQVYNFKRLVFTNCNYNLTKLLSLFSVNDFTCS 
                 
                 
                   QISPAAIASNCYSSLILDYFSYPLSMKSDLSVSSAGPISQFNYKQSFSNPTCLILATVPHNLT 
                 
                 
                   TITKPLKYSYINKCSRLLSDDRTEVPQLVNANQYSPCVSIVPSTVWEDGDYYRKQLSPLEG 
                 
                 
                   GGWLVASGSTVAMTEQLQMGFGITVQYGTDTNSVCPKLEFANDTKIASQLGNCVEYSLY 
                 
                 
                   GVSGRGVFQNCTAVGVRQQRFVYDAYQNLVGYYSDDGNYYCLRACVSVPVSVIYDKET 
                 
                 
                   KTHATLFGSVACEHISSTMSQYSRSTRSMLKRRDSTYGPLQTPVGCVLGLVNSSLFVEDC 
                 
                 
                   KLPLGQSLCALPDTPSTLTPRSVRSVPGEMRLASIAFNHPIQVDQLNSSYFKLSIPTNFSFG 
                 
                 
                   VTQEYIQTTIQKVTVDCKQYVCNGFQKCEQLLREYGQFCSKINQALHGANLRQDDSVRN 
                 
                 
                   LFASVKSSQSSPIIPGFGGDFNLTLLEPVSISTGSRSARSAIEDLLFDKVTIADPGYMQGYDD 
                 
                 
                   CMQQGPASARDLICAQYVAGYKVLPPLMDVNMEAAYTSSLLGSIAGVGWTAGLSSFAAI 
                 
                 
                   PFAQSIFYRLNGVGITQQVLSENQKLIANKFNQALGAMQTGFTTTNEAFQKVQDAVNNN 
                 
                 
                   AQALSKLASELSNTFGAISASIGDIIQRLDVLEQDAQIDRLINGRLTTLNAFVAQQLVRSES 
                 
                 
                   AALSAQLAKDKVNECVKAQSKRSGFCGQGTHIVSFVVNAPNGLYFMHVGYYPSNHIEV 
                 
                 
                   VSAYGLCDAANPTNCIAPVNGYFIKTNNTRIVDEWSYTGSSFYAPEPITSLNTKYVAPQVT 
                 
                 
                   YQNISTNLPPPLLGNSTGIDFQDELDEFFKNVSTSIPNFGSLTQINTTLLDLTYEMLSLQQV 
                 
                 
                   VKALNESYIDLKELGNYTYYNKWPWYIWLGFIAGLVALALCVFFILCCTGCGTNCMGKL 
                 
                 
                   KCNRCCDRYEEYDLEPHKVHVH 
                 
                 
                     
                 
                 
                   SEQ ID NO: 11 is a MERS-CoV Spike Glycoprotein, Nucleotide Sequence 
                 
                 
                   (EMC/2012, Genbank Ref: JX869059.2) 
                 
                 
                   ATGATACACTCAGTGTTTCTACTGATGTTCTTGTTAACACCTACAGAAAGTTACGTTG 
                 
                 
                   ATGTAGGGCCAGATTCTGTTAAGTCTGCTTGTATTGAGGTTGATATACAACAGACTTT 
                 
                 
                   CTTTGATAAAACTTGGCCTAGGCCAATTGATGTTTCTAAGGCTGACGGTATTATATAC 
                 
                 
                   CCTCAAGGCCGTACATATTCTAACATAACTATCACTTATCAAGGTCTTTTTCCCTATCA 
                 
                 
                   GGGAGACCATGGTGATATGTATGTTTACTCTGCAGGACATGCTACAGGCACAACTCC 
                 
                 
                   ACAAAAGTTGTTTGTAGCTAACTATTCTCAGGACGTCAAACAGTTTGCTAATGGGTTT 
                 
                 
                   GTCGTCCGTATAGGAGCAGCTGCCAATTCCACTGGCACTGTTATTATTAGCCCATCTA 
                 
                 
                   CCAGCGCTACTATACGAAAAATTTACCCTGCTTTTATGCTGGGTTCTTCAGTTGGTAA 
                 
                 
                   TTTCTCAGATGGTAAAATGGGCCGCTTCTTCAATCATACTCTAGTTCTTTTGCCCGATG 
                 
                 
                   GATGTGGCACTTTACTTAGAGCTTTTTATTGTATTCTAGAGCCTCGCTCTGGAAATCAT 
                 
                 
                   TGTCCTGCTGGCAATTCCTATACTTCTTTTGCCACTTATCACACTCCTGCAACAGATTG 
                 
                 
                   TTCTGATGGCAATTACAATCGTAATGCCAGTCTGAACTCTTTTAAGGAGTATTTTAAT 
                 
                 
                   TTACGTAACTGCACCTTTATGTACACTTATAACATTACCGAAGATGAGATTTTAGAGT 
                 
                 
                   GGTTTGGCATTACACAAACTGCTCAAGGTGTTCACCTCTTCTCATCTCGGTATGTTGA 
                 
                 
                   TTTGTACGGCGGCAATATGTTTCAATTTGCCACCTTGCCTGTTTATGATACTATTAAGT 
                 
                 
                   ATTATTCTATCATTCCTCACAGTATTCGTTCTATCCAAAGTGATAGAAAAGCTTGGGC 
                 
                 
                   TGCCTTCTACGTATATAAACTTCAACCGTTAACTTTCCTGTTGGATTTTTCTGTTGATG 
                 
                 
                   GTTATATACGCAGAGCTATAGACTGTGGTTTTAATGATTTGTCACAACTCCACTGCTC 
                 
                 
                   ATATGAATCCTTCGATGTTGAATCTGGAGTTTATTCAGTTTCGTCTTTCGAAGCAAAA 
                 
                 
                   CCTTCTGGCTCAGTTGTGGAACAGGCTGAAGGTGTTGAATGTGATTTTTCACCTCTTC 
                 
                 
                   TGTCTGGCACACCTCCTCAGGTTTATAATTTCAAGCGTTTGGTTTTTACCAATTGCAAT 
                 
                 
                   TATAATCTTACCAAATTGCTTTCACTTTTTTCTGTGAATGATTTTACTTGTAGTCAAAT 
                 
                 
                   ATCTCCAGCAGCAATTGCTAGCAACTGTTATTCTTCACTGATTTTGGATTACTTTTCAT 
                 
                 
                   ACCCACTTAGTATGAAATCCGATCTCAGTGTTAGTTCTGCTGGTCCAATATCCCAGTT 
                 
                 
                   TAATTATAAACAGTCCTTTTCTAATCCCACATGTTTGATTTTAGCGACTGTTCCTCATA 
                 
                 
                   ACCTTACTACTATTACTAAGCCTCTTAAGTACAGCTATATTAACAAGTGCTCTCGTCTT 
                 
                 
                   CTTTCTGATGATCGTACTGAAGTACCTCAGTTAGTGAACGCTAATCAATACTCACCCT 
                 
                 
                   GTGTATCCATTGTCCCATCCACTGTGTGGGAAGACGGTGATTATTATAGGAAACAACT 
                 
                 
                   ATCTCCACTTGAAGGTGGTGGCTGGCTTGTTGCTAGTGGCTCAACTGTTGCCATGACT 
                 
                 
                   GAGCAATTACAGATGGGCTTTGGTATTACAGTTCAATATGGTACAGACACCAATAGT 
                 
                 
                   GTTTGCCCCAAGCTTGAATTTGCTAATGACACAAAAATTGCCTCTCAATTAGGCAATT 
                 
                 
                   GCGTGGAATATTCCCTCTATGGTGTTTCGGGCCGTGGTGTTTTTCAGAATTGCACAGC 
                 
                 
                   TGTAGGTGTTCGACAGCAGCGCTTTGTTTATGATGCGTACCAGAATTTAGTTGGCTAT 
                 
                 
                   TATTCTGATGATGGCAACTACTACTGTTTGCGTGCTTGTGTTAGTGTTCCTGTTTCTGT 
                 
                 
                   CATCTATGATAAAGAAACTAAAACCCACGCTACTCTATTTGGTAGTGTTGCATGTGAA 
                 
                 
                   CACATTTCTTCTACCATGTCTCAATACTCCCGTTCTACGCGATCAATGCTTAAACGGC 
                 
                 
                   GAGATTCTACATATGGCCCCCTTCAGACACCTGTTGGTTGTGTCCTAGGACTTGTTAA 
                 
                 
                   TTCCTCTTTGTTCGTAGAGGACTGCAAGTTGCCTCTTGGTCAATCTCTCTGTGCTCTTC 
                 
                 
                   CTGACACACCTAGTACTCTCACACCTCGCAGTGTGCGCTCTGTTCCAGGTGAAATGCG 
                 
                 
                   CTTGGCATCCATTGCTTTTAATCATCCTATTCAGGTTGATCAACTTAATAGTAGTTATT 
                 
                 
                   TTAAATTAAGTATACCCACTAATTTTTCCTTTGGTGTGACTCAGGAGTACATTCAGAC 
                 
                 
                   AACCATTCAGAAAGTTACTGTTGATTGTAAACAGTACGTTTGCAATGGTTTCCAGAAG 
                 
                 
                   TGTGAGCAATTACTGCGCGAGTATGGCCAGTTTTGTTCCAAAATAAACCAGGCTCTCC 
                 
                 
                   ATGGTGCCAATTTACGCCAGGATGATTCTGTACGTAATTTGTTTGCGAGCGTGAAAAG 
                 
                 
                   CTCTCAATCATCTCCTATCATACCAGGTTTTGGAGGTGACTTTAATTTGACACTTCTAG 
                 
                 
                   AACCTGTTTCTATATCTACTGGCAGTCGTAGTGCACGTAGTGCTATTGAGGATTTGCT 
                 
                 
                   ATTTGACAAAGTCACTATAGCTGATCCTGGTTATATGCAAGGTTACGATGATTGCATG 
                 
                 
                   CAGCAAGGTCCAGCATCAGCTCGTGATCTTATTTGTGCTCAATATGTGGCTGGTTACA 
                 
                 
                   AAGTATTACCTCCTCTTATGGATGTTAATATGGAAGCCGCGTATACTTCATCTTTGCTT 
                 
                 
                   GGCAGCATAGCAGGTGTTGGCTGGACTGCTGGCTTATCCTCCTTTGCTGCTATTCCAT 
                 
                 
                   TTGCACAGAGTATCTTTTATAGGTTAAACGGTGTTGGCATTACTCAACAGGTTCTTTC 
                 
                 
                   AGAGAACCAAAAGCTTATTGCCAATAAGTTTAATCAGGCTCTGGGAGCTATGCAAAC 
                 
                 
                   AGGCTTCACTACAACTAATGAAGCTTTTCAGAAGGTTCAGGATGCTGTGAACAACAA 
                 
                 
                   TGCACAGGCTCTATCCAAATTAGCTAGCGAGCTATCTAATACTTTTGGTGCTATTTCC 
                 
                 
                   GCCTCTATTGGAGACATCATACAACGTCTTGATGTTCTCGAACAGGACGCCCAAATA 
                 
                 
                   GACAGACTTATTAATGGCCGTTTGACAACACTAAATGCTTTTGTTGCACAGCAGCTTG 
                 
                 
                   TTCGTTCCGAATCAGCTGCTCTTTCCGCTCAATTGGCTAAAGATAAAGTCAATGAGTG 
                 
                 
                   TGTCAAGGCACAATCCAAGCGTTCTGGATTTTGCGGTCAAGGCACACATATAGTGTCC 
                 
                 
                   TTTGTTGTAAATGCCCCTAATGGCCTTTACTTCATGCATGTTGGTTATTACCCTAGCAA 
                 
                 
                   CCACATTGAGGTTGTTTCTGCTTATGGTCTTTGCGATGCAGCTAACCCTACTAATTGTA 
                 
                 
                   TAGCCCCTGTTAATGGCTACTTTATTAAAACTAATAACACTAGGATTGTTGATGAGTG 
                 
                 
                   GTCATATACTGGCTCGTCCTTCTATGCACCTGAGCCCATTACCTCCCTTAATACTAAGT 
                 
                 
                   ATGTTGCACCACAGGTGACATACCAAAACATTTCTACTAACCTCCCTCCTCCTCTTCT 
                 
                 
                   CGGCAATTCCACCGGGATTGACTTCCAAGATGAGTTGGATGAGTTTTTCAAAAATGTT 
                 
                 
                   AGCACCAGTATACCTAATTTTGGTTCCCTAACACAGATTAATACTACATTACTCGATC 
                 
                 
                   TTACCTACGAGATGTTGTCTCTTCAACAAGTTGTTAAAGCCCTTAATGAGTCTTACAT 
                 
                 
                   AGACCTTAAAGAGCTTGGCAATTATACTTATTACAACAAATGGCCGTGGTACATTTGG 
                 
                 
                   CTTGGTTTCATTGCTGGGCTTGTTGCCTTAGCTCTATGCGTCTTCTTCATACTGTGCTG 
                 
                 
                   CACTGGTTGTGGCACAAACTGTATGGGAAAACTTAAGTGTAATCGTTGTTGTGATAG 
                 
                 
                   ATACGAGGAATACGACCTCGAGCCGCATAAGGTTCATGTTCACTAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 12 is a MERS-CoV Spike Glycoprotein, Nucleotide Sequence, 
                 
                 
                   Codon Optimized For Expression In Human Cells 
                 
                 
                   ATGATCCACAGCGTGTTCCTGCTGATGTTTCTGCTGACACCTACCGAGTCCTACGTGG 
                 
                 
                   ATGTGGGCCCAGACTCTGTGAAGAGCGCCTGCATCGAGGTGGACATCCAGCAGACAT 
                 
                 
                   TCTTTGACAAGACCTGGCCCAGACCCATCGACGTGAGCAAGGCAGACGGAATCATCT 
                 
                 
                   ACCCACAGGGACGCACATATAGCAACATCACAATCACCTACCAGGGCCTGTTCCCTT 
                 
                 
                   ATCAGGGCGACCACGGCGATATGTACGTGTATAGCGCCGGCCACGCAACCGGCACCA 
                 
                 
                   CACCACAGAAGCTGTTTGTGGCCAATTATTCCCAGGACGTGAAGCAGTTCGCCAACG 
                 
                 
                   GATTTGTGGTGCGGATCGGGGCCGCCGCCAACAGCACAGGCACCGTGATCATCTCTC 
                 
                 
                   CCAGCACATCCGCCACCATCAGAAAGATCTACCCTGCCTTTATGCTGGGCAGCTCCGT 
                 
                 
                   GGGCAACTTCTCCGATGGCAAGATGGGCAGGTTCTTTAATCACACACTGGTGCTGCTG 
                 
                 
                   CCAGACGGATGCGGCACCCTGCTGAGGGCCTTCTACTGTATCCTGGAGCCCCGCTCTG 
                 
                 
                   GAAATCACTGCCCTGCCGGCAACTCCTACACCTCTTTTGCCACATATCACACCCCTGC 
                 
                 
                   CACAGACTGTTCCGATGGCAATTATAACCGGAATGCCAGCCTGAACTCCTTCAAGGA 
                 
                 
                   GTACTTTAATCTGAGAAACTGCACCTTCATGTACACATATAATATCACCGAGGATGAG 
                 
                 
                   ATCCTGGAGTGGTTCGGCATCACACAGACCGCCCAGGGCGTGCACCTGTTTTCTAGCA 
                 
                 
                   GATACGTGGATCTGTATGGCGGCAACATGTTCCAGTTTGCCACACTGCCAGTGTATGA 
                 
                 
                   CACCATCAAGTACTATAGCATCATCCCCCACTCTATCCGGAGCATCCAGTCCGACAGA 
                 
                 
                   AAGGCCTGGGCCGCCTTCTACGTGTATAAGCTGCAGCCCCTGACCTTCCTGCTGGATT 
                 
                 
                   TTTCCGTGGACGGCTACATCCGGAGAGCCATCGATTGCGGCTTTAACGACCTGTCTCA 
                 
                 
                   GCTCCACTGTTCTTATGAGAGCTTCGATGTGGAGTCTGGCGTGTACAGCGTGTCCTCT 
                 
                 
                   TTTGAGGCCAAGCCATCTGGCAGCGTGGTGGAGCAGGCAGAGGGAGTGGAGTGCGA 
                 
                 
                   CTTCTCCCCACTGCTGTCTGGCACACCACCTCAGGTGTATAATTTCAAGAGGCTGGTG 
                 
                 
                   TTTACAAACTGTAATTACAACCTGACCAAGCTGCTGTCCCTGTTCTCTGTGAACGACT 
                 
                 
                   TTACCTGCAGCCAGATCTCCCCTGCCGCCATCGCCTCCAATTGTTATAGCTCCCTGAT 
                 
                 
                   CCTGGATTACTTCTCTTATCCCCTGTCTATGAAGAGCGACCTGTCCGTGTCTAGCGCC 
                 
                 
                   GGCCCTATCAGCCAGTTTAATTACAAGCAGTCCTTCTCTAACCCCACATGCCTGATCC 
                 
                 
                   TGGCCACCGTGCCTCACAACCTGACCACAATCACAAAGCCACTGAAGTACTCCTATA 
                 
                 
                   TCAATAAGTGCAGCAGGCTGCTGTCCGACGATCGCACCGAGGTGCCTCAGCTCGTGA 
                 
                 
                   ACGCCAACCAGTACTCTCCATGCGTGAGCATCGTGCCATCCACCGTGTGGGAGGACG 
                 
                 
                   GCGATTACTATAGAAAGCAGCTCAGCCCACTGGAGGGAGGAGGATGGCTGGTGGCC 
                 
                 
                   AGCGGCTCCACAGTGGCCATGACCGAGCAGCTCCAGATGGGCTTCGGCATCACAGTG 
                 
                 
                   CAGTACGGCACAGATACCAATAGCGTGTGCCCCAAGCTGGAGTTTGCCAACGACACC 
                 
                 
                   AAGATCGCCTCCCAGCTCGGCAATTGCGTGGAGTACTCCCTGTATGGCGTGTCTGGCA 
                 
                 
                   GAGGCGTGTTCCAGAACTGTACAGCCGTGGGCGTGCGGCAGCAGCGGTTCGTGTACG 
                 
                 
                   ATGCCTATCAGAACCTGGTGGGCTACTATAGCGACGATGGCAATTACTATTGCCTGA 
                 
                 
                   GGGCATGCGTGAGCGTGCCCGTGAGCGTGATCTACGACAAGGAGACAAAGACCCAC 
                 
                 
                   GCCACCCTGTTCGGCTCCGTGGCCTGCGAGCACATCTCCTCTACAATGTCTCAGTATT 
                 
                 
                   CTAGGAGCACCCGCTCTATGCTGAAGAGGCGCGACAGCACATACGGACCACTGCAGA 
                 
                 
                   CCCCTGTGGGATGCGTGCTGGGCCTGGTGAACAGCAGCCTGTTTGTGGAGGATTGCA 
                 
                 
                   AGCTGCCACTGGGCCAGTCTCTGTGCGCACTGCCAGACACCCCCAGCACACTGACCC 
                 
                 
                   CACGGTCTGTGAGAAGCGTGCCCGGAGAGATGAGACTGGCCAGCATCGCCTTCAATC 
                 
                 
                   ACCCTATCCAGGTGGATCAGCTCAACAGCAGCTACTTTAAGCTGAGCATCCCAACAA 
                 
                 
                   ACTTCTCCTTTGGCGTGACCCAGGAGTATATCCAGACCACAATCCAGAAGGTGACCG 
                 
                 
                   TGGACTGCAAGCAGTACGTGTGCAATGGCTTCCAGAAGTGCGAGCAGCTCCTGAGGG 
                 
                 
                   AGTATGGCCAGTTTTGTTCCAAGATCAATCAGGCCCTGCACGGAGCCAACCTGAGGC 
                 
                 
                   AGGACGATTCCGTGAGAAACCTGTTCGCCTCTGTGAAGTCCTCTCAGAGCTCCCCTAT 
                 
                 
                   CATCCCAGGCTTCGGCGGCGACTTCAACCTGACCCTGCTGGAGCCCGTGTCCATCTCT 
                 
                 
                   ACCGGCAGCAGGTCCGCCCGCAGCGCCATCGAGGATCTGCTGTTTGACAAGGTGACC 
                 
                 
                   ATCGCCGACCCAGGCTACATGCAGGGCTATGACGATTGCATGCAGCAGGGACCAGCC 
                 
                 
                   TCCGCCCGCGATCTGATCTGTGCCCAGTACGTGGCCGGCTATAAGGTGCTGCCACCCC 
                 
                 
                   TGATGGACGTGAACATGGAGGCCGCCTATACATCTAGCCTGCTGGGCAGCATCGCAG 
                 
                 
                   GAGTGGGATGGACCGCCGGCCTGTCCTCTTTCGCCGCAATCCCTTTTGCCCAGTCTAT 
                 
                 
                   CTTCTACCGGCTGAACGGCGTGGGCATCACACAGCAGGTGCTGAGCGAGAATCAGAA 
                 
                 
                   GCTGATCGCCAATAAGTTCAACCAGGCCCTGGGGGCCATGCAGACCGGCTTTACCAC 
                 
                 
                   AACCAACGAGGCCTTCCAGAAGGTGCAGGATGCCGTGAACAATAACGCACAGGCCCT 
                 
                 
                   GTCCAAGCTGGCCTCCGAGCTGTCTAATACCTTCGGGGCCATCAGCGCCAGCATCGG 
                 
                 
                   CGACATCATCCAGCGCCTGGACGTGCTGGAGCAGGATGCCCAGATCGACAGGCTGAT 
                 
                 
                   CAATGGCCGCCTGACAACCCTGAACGCCTTTGTGGCACAGCAGCTCGTGCGGAGCGA 
                 
                 
                   GTCTGCCGCCCTGAGCGCCCAGCTCGCCAAGGACAAGGTGAACGAGTGCGTGAAGGC 
                 
                 
                   CCAGAGCAAGCGGTCCGGCTTTTGTGGCCAGGGCACCCACATCGTGTCCTTCGTGGTG 
                 
                 
                   AATGCCCCTAACGGCCTGTACTTTATGCATGTGGGCTACTATCCAAGCAACCACATCG 
                 
                 
                   AGGTGGTGTCCGCCTATGGCCTGTGCGATGCCGCCAATCCTACAAACTGTATCGCCCC 
                 
                 
                   AGTGAATGGCTACTTCATCAAGACCAATAACACACGGATCGTGGACGAGTGGTCCTA 
                 
                 
                   CACCGGCAGCTCCTTTTATGCCCCCGAGCCTATCACATCTCTGAACACCAAGTACGTG 
                 
                 
                   GCCCCACAGGTGACATATCAGAATATCAGCACCAACCTGCCTCCACCCCTGCTGGGC 
                 
                 
                   AATTCCACCGGCATCGACTTCCAGGATGAGCTGGACGAGTTCTTTAAGAATGTGAGC 
                 
                 
                   ACATCCATCCCCAACTTTGGCAGCCTGACCCAGATCAACACAACCCTGCTGGATCTGA 
                 
                 
                   CATACGAGATGCTGTCTCTGCAGCAGGTGGTGAAGGCCCTGAATGAGAGCTACATCG 
                 
                 
                   ACCTGAAGGAGCTGGGCAATTATACCTACTATAACAAGTGGCCTTGGTACATCTGGC 
                 
                 
                   TGGGCTTCATCGCAGGCCTGGTGGCCCTGGCCCTGTGCGTGTTCTTTATCCTGTGCTGT 
                 
                 
                   ACAGGCTGCGGCACCAATTGTATGGGCAAGCTGAAGTGTAACCGGTGCTGTGATAGA 
                 
                 
                   TACGAGGAGTATGACCTGGAGCCACACAAGGTGCATGTGCACTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 13 is a SARS-CoV-2 “Proline Modified” Spike Glycoprotein, 
                 
                 
                   Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLAL 
                 
                 
                   HRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTL 
                 
                 
                   KSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVAD 
                 
                 
                   YSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYK 
                 
                 
                   LPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVE 
                 
                 
                   GFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNF 
                 
                 
                   NGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTS 
                 
                 
                   NQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDI 
                 
                 
                   PIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILP 
                 
                 
                   VSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQI 
                 
                 
                   YKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC 
                 
                 
                   AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGV 
                 
                 
                   TQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFG 
                 
                 
                   AISSVLNDILSRLD PP EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECV 
                 
                 
                   LGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPRE 
                 
                 
                   GVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELD 
                 
                 
                   KYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWP 
                 
                 
                   WYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT 
                 
                 
                     
                 
                 
                   SEQ ID NO: 14 is a SARS-CoV-2 “Proline Modified” Spike Glycoprotein, 
                 
                 
                   Nucleotide Sequence 
                 
                 
                   ATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAAC 
                 
                 
                   CAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCT 
                 
                 
                   GACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTT 
                 
                 
                   TTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGG 
                 
                 
                   TTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTC 
                 
                 
                   TAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTA 
                 
                 
                   CTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAA 
                 
                 
                   TGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGA 
                 
                 
                   GTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTC 
                 
                 
                   TTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTA 
                 
                 
                   AGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCG 
                 
                 
                   TGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATT 
                 
                 
                   AACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTG 
                 
                 
                   ATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCT 
                 
                 
                   AGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGT 
                 
                 
                   GCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAA 
                 
                 
                   GGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTC 
                 
                 
                   CTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCT 
                 
                 
                   GTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTAT 
                 
                 
                   ATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAAT 
                 
                 
                   GATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCA 
                 
                 
                   GACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAG 
                 
                 
                   ATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGG 
                 
                 
                   TGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAG 
                 
                 
                   AGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAA 
                 
                 
                   GGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGG 
                 
                 
                   TTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACT 
                 
                 
                   GTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACT 
                 
                 
                   TCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTT 
                 
                 
                   CCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGAC 
                 
                 
                   ACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCA 
                 
                 
                   GGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAG 
                 
                 
                   TCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGT 
                 
                 
                   TCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACT 
                 
                 
                   CATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGAC 
                 
                 
                   TAATTCTCCTCGGCGGGCACGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATG 
                 
                 
                   TCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAA 
                 
                 
                   ATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGT 
                 
                 
                   AGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAA 
                 
                 
                   TATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAG 
                 
                 
                   ACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAA 
                 
                 
                   TTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAG 
                 
                 
                   CAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGC 
                 
                 
                   TTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTG 
                 
                 
                   CACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGC 
                 
                 
                   TCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCA 
                 
                 
                   GGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTG 
                 
                 
                   GAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATA 
                 
                 
                   GTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACT 
                 
                 
                   TCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAG 
                 
                 
                   CTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACCCTC 
                 
                 
                   CTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGA 
                 
                 
                   CATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGC 
                 
                 
                   TGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGA 
                 
                 
                   AAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCA 
                 
                 
                   TGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCAT 
                 
                 
                   GATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGT 
                 
                 
                   TTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGT 
                 
                 
                   GTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTG 
                 
                 
                   CAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACA 
                 
                 
                   TCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTC 
                 
                 
                   AAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCG 
                 
                 
                   ATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATGGCCATGGTACATTTGGC 
                 
                 
                   TAGGTTTTATAGCTGGCTTGATTGCCATAGTAATGGTGACAATTATGCTTTGCTGTAT 
                 
                 
                   GACCAGTTGCTGTAGTTGTCTCAAGGGCTGTTGTTCTTGTGGATCCTGCTGCAAATTT 
                 
                 
                   GATGAAGACGACTCTGAGCCAGTGCTCAAAGGAGTCAAATTACATTACACATAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 15 is a SARS-CoV-2 “Proline Modified” Spike Glycoprotein, 
                 
                 
                   Nucleotide Sequence, Codon Optimized for Expression In Human Cells 
                 
                 
                   ATGTTCGTGTTTCTGGTGCTGCTGCCTCTGGTGAGCTCCCAGTGCGTGAACCTGACCA 
                 
                 
                   CAAGGACCCAGCTCCCCCCTGCCTATACCAATTCCTTCACACGGGGCGTGTACTATCC 
                 
                 
                   AGACAAGGTGTTTAGATCTAGCGTGCTGCACTCCACACAGGATCTGTTTCTGCCCTTC 
                 
                 
                   TTTTCTAACGTGACCTGGTTCCACGCCATCCATGTGAGCGGCACCAATGGCACAAAGC 
                 
                 
                   GGTTCGACAATCCTGTGCTGCCCTTCAACGATGGCGTGTACTTCGCCTCCACCGAGAA 
                 
                 
                   GTCTAACATCATCAGAGGCTGGATCTTTGGCACCACACTGGACAGCAAGACACAGTC 
                 
                 
                   CCTGCTGATCGTGAACAATGCCACCAACGTGGTCATCAAGGTGTGCGAGTTCCAGTTT 
                 
                 
                   TGTAATGATCCTTTCCTGGGCGTGTACTATCACAAGAACAATAAGTCTTGGATGGAGA 
                 
                 
                   GCGAGTTTCGCGTGTATTCCTCTGCCAACAATTGCACATTTGAGTACGTGTCCCAGCC 
                 
                 
                   ATTCCTGATGGACCTGGAGGGCAAGCAGGGCAATTTCAAGAACCTGAGGGAGTTCGT 
                 
                 
                   GTTTAAGAATATCGATGGCTACTTCAAGATCTACTCCAAGCACACCCCTATCAACCTG 
                 
                 
                   GTGCGCGACCTGCCACAGGGCTTCTCTGCCCTGGAGCCTCTGGTGGATCTGCCAATCG 
                 
                 
                   GCATCAACATCACCCGGTTTCAGACACTGCTGGCCCTGCACAGAAGCTACCTGACAC 
                 
                 
                   CTGGCGACAGCTCCTCTGGATGGACCGCCGGGGCCGCCGCCTACTATGTGGGCTATCT 
                 
                 
                   GCAGCCAAGGACCTTCCTGCTGAAGTACAACGAGAATGGCACCATCACAGACGCAGT 
                 
                 
                   GGATTGCGCCCTGGACCCCCTGTCTGAGACCAAGTGTACACTGAAGAGCTTTACCGT 
                 
                 
                   GGAGAAGGGCATCTATCAGACAAGCAATTTCAGGGTGCAGCCCACCGAGTCCATCGT 
                 
                 
                   GCGCTTTCCAAATATCACAAACCTGTGCCCCTTTGGCGAGGTGTTCAACGCAACCAGG 
                 
                 
                   TTCGCCAGCGTGTACGCATGGAATAGGAAGCGCATCTCCAACTGCGTGGCCGACTAT 
                 
                 
                   TCTGTGCTGTACAACAGCGCCTCCTTCTCTACCTTTAAGTGCTATGGCGTGAGCCCCA 
                 
                 
                   CAAAGCTGAATGACCTGTGCTTTACCAACGTGTACGCCGATTCCTTCGTGATCAGGGG 
                 
                 
                   CGACGAGGTGCGCCAGATCGCACCAGGACAGACAGGCAAGATCGCCGACTACAATT 
                 
                 
                   ATAAGCTGCCCGACGATTTCACCGGCTGCGTGATCGCCTGGAACTCTAACAATCTGG 
                 
                 
                   ATAGCAAAGTGGGCGGCAACTACAATTATCTGTACCGGCTGTTTAGAAAGTCTAATC 
                 
                 
                   TGAAGCCTTTCGAGAGGGACATCTCCACAGAGATCTACCAGGCCGGCTCTACCCCAT 
                 
                 
                   GCAATGGCGTGGAGGGCTTTAACTGTTATTTCCCCCTGCAGAGCTACGGCTTCCAGCC 
                 
                 
                   TACAAACGGCGTGGGCTATCAGCCATACCGCGTGGTGGTGCTGTCTTTTGAGCTGCTG 
                 
                 
                   CACGCACCAGCAACAGTGTGCGGACCTAAGAAGAGCACCAATCTGGTGAAGAACAA 
                 
                 
                   GTGCGTGAACTTCAACTTCAACGGCCTGACCGGCACAGGCGTGCTGACCGAGTCCAA 
                 
                 
                   CAAGAAGTTCCTGCCCTTTCAGCAGTTCGGCAGGGACATCGCAGATACCACAGACGC 
                 
                 
                   CGTGCGCGACCCCCAGACCCTGGAGATCCTGGACATCACACCTTGCTCCTTCGGCGGC 
                 
                 
                   GTGTCTGTGATCACACCTGGCACCAATACAAGCAACCAGGTGGCCGTGCTGTATCAG 
                 
                 
                   GACGTGAATTGTACCGAGGTGCCAGTGGCCATCCACGCCGATCAGCTCACCCCCACA 
                 
                 
                   TGGCGGGTGTACTCTACCGGCAGCAACGTGTTCCAGACAAGAGCCGGCTGCCTGATC 
                 
                 
                   GGAGCCGAGCATGTGAACAATAGCTATGAGTGCGACATCCCCATCGGAGCCGGCATC 
                 
                 
                   TGTGCCTCCTACCAGACCCAGACAAACTCCCCTCGGAGAGCCCGGTCTGTGGCCAGC 
                 
                 
                   CAGTCCATCATCGCCTATACCATGAGCCTGGGGGCCGAGAACAGCGTGGCCTACTCC 
                 
                 
                   AACAATTCTATCGCCATCCCCACCAACTTCACAATCTCCGTGACCACAGAGATCCTGC 
                 
                 
                   CTGTGAGCATGACCAAGACATCCGTGGACTGCACAATGTATATCTGTGGCGATTCCA 
                 
                 
                   CCGAGTGCTCTAACCTGCTGCTGCAGTACGGCTCTTTTTGTACCCAGCTCAACAGAGC 
                 
                 
                   CCTGACAGGCATCGCCGTGGAGCAGGACAAGAACACACAGGAGGTGTTCGCCCAGG 
                 
                 
                   TGAAGCAGATCTACAAGACCCCACCCATCAAGGACTTTGGCGGCTTCAACTTCAGCC 
                 
                 
                   AGATCCTGCCAGATCCCAGCAAGCCTTCCAAGCGGTCTTTTATCGAGGACCTGCTGTT 
                 
                 
                   CAACAAGGTGACCCTGGCCGATGCCGGCTTCATCAAGCAGTATGGCGATTGCCTGGG 
                 
                 
                   CGACATCGCCGCCAGAGACCTGATCTGTGCCCAGAAGTTTAATGGCCTGACCGTGCT 
                 
                 
                   GCCTCCACTGCTGACAGATGAGATGATCGCCCAGTACACATCTGCCCTGCTGGCCGG 
                 
                 
                   CACCATCACAAGCGGATGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCATTTGC 
                 
                 
                   CATGCAGATGGCCTATCGGTTCAACGGCATCGGCGTGACCCAGAATGTGCTGTACGA 
                 
                 
                   GAACCAGAAGCTGATCGCCAATCAGTTTAACTCCGCCATCGGCAAGATCCAGGACTC 
                 
                 
                   TCTGAGCTCCACAGCCAGCGCCCTGGGCAAGCTGCAGGATGTGGTGAATCAGAACGC 
                 
                 
                   CCAGGCCCTGAATACCCTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAGCAG 
                 
                 
                   CGTGCTGAACGACATCCTGAGCCGGCTGGACCCCCCTGAGGCAGAGGTGCAGATCGA 
                 
                 
                   CCGGCTGATCACAGGCAGACTGCAGTCCCTGCAGACCTACGTGACACAGCAGCTCAT 
                 
                 
                   CAGGGCCGCCGAGATCAGGGCCTCTGCCAATCTGGCCGCCACCAAGATGAGCGAGTG 
                 
                 
                   CGTGCTGGGCCAGTCCAAGAGAGTGGACTTTTGTGGCAAGGGCTATCACCTGATGAG 
                 
                 
                   CTTCCCACAGTCCGCCCCCCACGGAGTGGTGTTTCTGCATGTGACCTACGTGCCTGCC 
                 
                 
                   CAGGAGAAGAACTTCACCACAGCCCCAGCCATCTGCCACGATGGCAAGGCCCACTTT 
                 
                 
                   CCCAGGGAGGGCGTGTTCGTGAGCAACGGCACCCACTGGTTTGTGACACAGCGCAAT 
                 
                 
                   TTCTACGAGCCTCAGATCATCACCACAGACAATACATTCGTGTCCGGCAACTGTGACG 
                 
                 
                   TGGTCATCGGCATCGTGAACAATACCGTGTATGATCCCCTGCAGCCTGAGCTGGACTC 
                 
                 
                   TTTTAAGGAGGAGCTGGATAAGTACTTCAAGAATCACACCAGCCCCGACGTGGATCT 
                 
                 
                   GGGCGACATCTCTGGCATCAATGCCAGCGTGGTGAACATCCAGAAGGAGATCGACAG 
                 
                 
                   GCTGAACGAGGTGGCCAAGAATCTGAACGAGTCCCTGATCGATCTGCAGGAGCTGGG 
                 
                 
                   CAAGTATGAGCAGTACATCAAGTGGCCATGGTATATCTGGCTGGGCTTCATCGCCGG 
                 
                 
                   CCTGATCGCCATCGTGATGGTGACCATCATGCTGTGCTGTATGACAAGCTGCTGTTCC 
                 
                 
                   TGCCTGAAGGGCTGCTGTTCTTGTGGCTCCTGCTGTAAGTTTGATGAGGACGATAGCG 
                 
                 
                   AGCCCGTGCTGAAGGGCGTGAAGCTGCACTACACCTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 16 is a SARS-CoV-2 “Furin Cleavage Modified” Spike 
                 
                 
                   Glycoprotein, Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLAL 
                 
                 
                   HRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTL 
                 
                 
                   KSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVAD 
                 
                 
                   YSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYK 
                 
                 
                   LPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVE 
                 
                 
                   GFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNF 
                 
                 
                   NGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTS 
                 
                 
                   NQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDI 
                 
                 
                   PIGAGICASYQTQTNSP GS A S SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILP 
                 
                 
                   VSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQI 
                 
                 
                   YKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC 
                 
                 
                   AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGV 
                 
                 
                   TQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFG 
                 
                 
                   AISSVLNDILSRLD KV EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSEC 
                 
                 
                   VLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPR 
                 
                 
                   EGVEVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEEL 
                 
                 
                   DKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWP 
                 
                 
                   WYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT 
                 
                 
                     
                 
                 
                   SEQ ID NO: 17 is a SARS-CoV-2 “Furin Cleavage Modified” Spike 
                 
                 
                   Glycoprotein, Nucleotide Sequence 
                 
                 
                   ATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAAC 
                 
                 
                   CAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCT 
                 
                 
                   GACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTT 
                 
                 
                   TTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGG 
                 
                 
                   TTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTC 
                 
                 
                   TAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTA 
                 
                 
                   CTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAA 
                 
                 
                   TGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGA 
                 
                 
                   GTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTC 
                 
                 
                   TTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTA 
                 
                 
                   AGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCG 
                 
                 
                   TGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATT 
                 
                 
                   AACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTG 
                 
                 
                   ATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCT 
                 
                 
                   AGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGT 
                 
                 
                   GCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAA 
                 
                 
                   GGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTC 
                 
                 
                   CTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCT 
                 
                 
                   GTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTAT 
                 
                 
                   ATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAAT 
                 
                 
                   GATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCA 
                 
                 
                   GACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAG 
                 
                 
                   ATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGG 
                 
                 
                   TGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAG 
                 
                 
                   AGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAA 
                 
                 
                   GGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGG 
                 
                 
                   TTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACT 
                 
                 
                   GTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACT 
                 
                 
                   TCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTT 
                 
                 
                   CCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGAC 
                 
                 
                   ACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCA 
                 
                 
                   GGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAG 
                 
                 
                   TCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGT 
                 
                 
                   TCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACT 
                 
                 
                   CATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGAC 
                 
                 
                   TAATTCTCCTGGTAGTGCAAGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATG 
                 
                 
                   TCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAA 
                 
                 
                   ATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGT 
                 
                 
                   AGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAA 
                 
                 
                   TATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAG 
                 
                 
                   ACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAA 
                 
                 
                   TTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAG 
                 
                 
                   CAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGC 
                 
                 
                   TTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTG 
                 
                 
                   CACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGC 
                 
                 
                   TCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCA 
                 
                 
                   GGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTG 
                 
                 
                   GAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATA 
                 
                 
                   GTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACT 
                 
                 
                   TCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAG 
                 
                 
                   CTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACAAAG 
                 
                 
                   TTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGA 
                 
                 
                   CATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGC 
                 
                 
                   TGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGA 
                 
                 
                   AAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCA 
                 
                 
                   TGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCAT 
                 
                 
                   GATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGT 
                 
                 
                   TTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGT 
                 
                 
                   GTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTG 
                 
                 
                   CAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACA 
                 
                 
                   TCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTC 
                 
                 
                   AAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCG 
                 
                 
                   ATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATGGCCATGGTACATTTGGC 
                 
                 
                   TAGGTTTTATAGCTGGCTTGATTGCCATAGTAATGGTGACAATTATGCTTTGCTGTAT 
                 
                 
                   GACCAGTTGCTGTAGTTGTCTCAAGGGCTGTTGTTCTTGTGGATCCTGCTGCAAATTT 
                 
                 
                   GATGAAGACGACTCTGAGCCAGTGCTCAAAGGAGTCAAATTACATTACACATAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 18 is a SARS-CoV-2 “Furin Cleavage Modified” Spike 
                 
                 
                   Glycoprotein, Nucleotide Sequence, Codon Optimized For Expression In 
                 
                 
                   Human Cells 
                 
                 
                   ATGTTCGTGTTTCTGGTGCTGCTGCCTCTGGTGAGCTCCCAGTGCGTGAACCTGACCA 
                 
                 
                   CAAGGACCCAGCTCCCCCCTGCCTATACCAATTCCTTCACAAGGGGCGTGTACTATCC 
                 
                 
                   CGACAAGGTGTTTCGCTCTAGCGTGCTGCACAGCACACAGGATCTGTTTCTGCCTTTC 
                 
                 
                   TTTTCCAACGTGACCTGGTTCCACGCCATCCATGTGAGCGGCACCAATGGCACAAAG 
                 
                 
                   AGGTTCGACAATCCAGTGCTGCCCTTTAACGATGGCGTGTACTTCGCCTCTACCGAGA 
                 
                 
                   AGAGCAACATCATCCGCGGCTGGATCTTTGGCACCACACTGGACTCCAAGACACAGT 
                 
                 
                   CTCTGCTGATCGTGAACAATGCCACCAACGTGGTCATCAAGGTGTGCGAGTTCCAGTT 
                 
                 
                   TTGTAATGATCCATTCCTGGGCGTGTACTATCACAAGAACAATAAGAGCTGGATGGA 
                 
                 
                   GTCCGAGTTTCGCGTGTATTCCTCTGCCAACAATTGCACATTTGAGTACGTGTCCCAG 
                 
                 
                   CCCTTCCTGATGGACCTGGAGGGCAAGCAGGGCAATTTCAAGAACCTGCGGGAGTTC 
                 
                 
                   GTGTTTAAGAATATCGATGGCTACTTCAAGATCTACAGCAAGCACACCCCAATCAAC 
                 
                 
                   CTGGTGAGAGACCTGCCACAGGGCTTCTCCGCCCTGGAGCCACTGGTGGATCTGCCC 
                 
                 
                   ATCGGCATCAACATCACCAGGTTTCAGACACTGCTGGCCCTGCACCGCAGCTACCTG 
                 
                 
                   ACACCAGGCGACAGCTCCTCTGGATGGACCGCCGGGGCCGCCGCCTACTATGTGGGC 
                 
                 
                   TATCTGCAGCCCCGGACCTTCCTGCTGAAGTACAACGAGAATGGCACCATCACAGAC 
                 
                 
                   GCAGTGGATTGCGCCCTGGACCCCCTGTCCGAGACCAAGTGTACACTGAAGTCTTTTA 
                 
                 
                   CCGTGGAGAAGGGCATCTATCAGACATCTAATTTCCGGGTGCAGCCTACCGAGAGCA 
                 
                 
                   TCGTGAGATTTCCCAATATCACAAACCTGTGCCCTTTTGGCGAGGTGTTCAACGCCAC 
                 
                 
                   CAGATTCGCCAGCGTGTACGCCTGGAATCGGAAGAGAATCAGCAACTGCGTGGCCGA 
                 
                 
                   CTATTCCGTGCTGTACAACTCTGCCAGCTTCTCCACCTTTAAGTGCTATGGCGTGTCTC 
                 
                 
                   CCACAAAGCTGAATGACCTGTGCTTTACCAACGTGTACGCCGATAGCTTCGTGATCAG 
                 
                 
                   GGGCGACGAGGTGAGACAGATCGCACCAGGACAGACAGGCAAGATCGCAGACTACA 
                 
                 
                   ATTATAAGCTGCCTGACGATTTCACCGGCTGCGTGATCGCCTGGAACAGCAACAATCT 
                 
                 
                   GGATTCCAAAGTGGGCGGCAACTACAATTATCTGTACAGGCTGTTTCGCAAGTCCAA 
                 
                 
                   TCTGAAGCCATTCGAGCGGGACATCAGCACAGAGATCTACCAGGCAGGCTCCACCCC 
                 
                 
                   ATGCAATGGAGTGGAGGGCTTTAACTGTTATTTCCCTCTGCAGTCTTACGGCTTCCAG 
                 
                 
                   CCAACAAACGGCGTGGGCTATCAGCCCTACAGAGTGGTGGTGCTGTCCTTTGAGCTG 
                 
                 
                   CTGCACGCACCTGCAACAGTGTGCGGACCAAAGAAGTCTACCAATCTGGTGAAGAAC 
                 
                 
                   AAGTGCGTGAACTTCAACTTCAACGGCCTGACCGGCACAGGCGTGCTGACCGAGTCC 
                 
                 
                   AACAAGAAGTTCCTGCCTTTTCAGCAGTTCGGCAGAGACATCGCCGATACCACAGAC 
                 
                 
                   GCCGTGAGAGACCCTCAGACCCTGGAGATCCTGGACATCACACCATGCTCTTTCGGC 
                 
                 
                   GGCGTGAGCGTGATCACACCAGGCACCAATACAAGCAACCAGGTGGCCGTGCTGTAT 
                 
                 
                   CAGGACGTGAATTGTACCGAGGTGCCCGTGGCCATCCACGCAGATCAGCTCACCCCT 
                 
                 
                   ACATGGAGGGTGTACTCCACCGGCTCTAACGTGTTCCAGACACGCGCCGGATGCCTG 
                 
                 
                   ATCGGAGCCGAGCATGTGAACAATTCTTATGAGTGCGACATCCCTATCGGAGCCGGC 
                 
                 
                   ATCTGTGCCAGCTACCAGACCCAGACAAACAGCCCAGGCTCCGCCAGCTCCGTGGCC 
                 
                 
                   TCTCAGAGCATCATCGCCTATACCATGAGCCTGGGGGCCGAGAATAGCGTGGCCTAC 
                 
                 
                   TCTAACAATAGCATCGCCATCCCTACCAACTTCACAATCTCCGTGACCACAGAGATCC 
                 
                 
                   TGCCAGTGTCCATGACCAAGACATCTGTGGACTGCACAATGTATATCTGTGGCGATTC 
                 
                 
                   TACCGAGTGCAGCAACCTGCTGCTGCAGTACGGCAGCTTTTGTACCCAGCTCAACCG 
                 
                 
                   GGCCCTGACAGGAATCGCAGTGGAGCAGGACAAGAACACACAGGAGGTGTTCGCCC 
                 
                 
                   AGGTGAAGCAGATCTACAAGACCCCACCCATCAAGGACTTTGGCGGCTTCAACTTCA 
                 
                 
                   GCCAGATCCTGCCCGATCCTTCCAAGCCATCTAAGAGGAGCTTTATCGAGGACCTGCT 
                 
                 
                   GTTCAACAAGGTGACCCTGGCCGATGCCGGCTTCATCAAGCAGTATGGCGATTGCCT 
                 
                 
                   GGGCGACATCGCAGCCCGCGACCTGATCTGTGCCCAGAAGTTTAATGGCCTGACCGT 
                 
                 
                   GCTGCCTCCACTGCTGACAGATGAGATGATCGCACAGTACACATCCGCCCTGCTGGC 
                 
                 
                   CGGCACCATCACATCTGGATGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCCTTT 
                 
                 
                   GCCATGCAGATGGCCTATAGATTCAACGGCATCGGCGTGACCCAGAATGTGCTGTAC 
                 
                 
                   GAGAACCAGAAGCTGATCGCCAATCAGTTTAACTCCGCCATCGGCAAGATCCAGGAC 
                 
                 
                   TCCCTGTCTAGCACAGCCTCTGCCCTGGGCAAGCTGCAGGATGTGGTGAATCAGAAC 
                 
                 
                   GCCCAGGCCCTGAATACCCTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAGC 
                 
                 
                   AGCGTGCTGAACGACATCCTGAGCCGGCTGGACAAGGTGGAGGCAGAGGTGCAGAT 
                 
                 
                   CGACAGGCTGATCACAGGCCGCCTGCAGAGCCTGCAGACCTACGTGACACAGCAGCT 
                 
                 
                   CATCAGGGCCGCCGAGATCAGAGCCTCCGCCAATCTGGCCGCCACCAAGATGTCTGA 
                 
                 
                   GTGCGTGCTGGGCCAGAGCAAGCGCGTGGACTTTTGTGGCAAGGGCTATCACCTGAT 
                 
                 
                   GTCCTTCCCACAGTCTGCCCCTCACGGAGTGGTGTTTCTGCATGTGACCTACGTGCCA 
                 
                 
                   GCCCAGGAGAAGAACTTCACCACAGCCCCCGCAATCTGCCACGATGGCAAGGCACAC 
                 
                 
                   TTTCCTCGGGAGGGCGTGTTCGTGTCTAACGGCACCCACTGGTTTGTGACACAGAGAA 
                 
                 
                   ATTTCTACGAGCCACAGATCATCACCACAGACAATACATTCGTGAGCGGCAACTGTG 
                 
                 
                   ACGTGGTCATCGGCATCGTGAACAATACCGTGTATGATCCTCTGCAGCCAGAGCTGG 
                 
                 
                   ACTCCTTTAAGGAGGAGCTGGATAAGTACTTCAAGAATCACACCTCTCCCGACGTGG 
                 
                 
                   ATCTGGGCGACATCAGCGGCATCAATGCCTCCGTGGTGAACATCCAGAAGGAGATCG 
                 
                 
                   ACAGGCTGAACGAGGTGGCCAAGAATCTGAACGAGTCCCTGATCGATCTGCAGGAGC 
                 
                 
                   TGGGCAAGTATGAGCAGTACATCAAGTGGCCCTGGTATATCTGGCTGGGCTTCATCG 
                 
                 
                   CCGGCCTGATCGCCATCGTGATGGTGACCATCATGCTGTGCTGTATGACATCCTGCTG 
                 
                 
                   TTCTTGCCTGAAGGGCTGCTGTAGCTGTGGCTCCTGCTGTAAGTTTGATGAGGACGAT 
                 
                 
                   AGCGAGCCTGTGCTGAAGGGCGTGAAGCTGCACTACACCTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 19 is a SARS-CoV-2 “Proline &amp; Furin Cleavage Modified” Spike 
                 
                 
                   Glycoprotein, Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLAL 
                 
                 
                   HRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTL 
                 
                 
                   KSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVAD 
                 
                 
                   YSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYK 
                 
                 
                   LPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVE 
                 
                 
                   GFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNF 
                 
                 
                   NGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTS 
                 
                 
                   NQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDI 
                 
                 
                   PIGAGICASYQTQTNSP GS A S SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILP 
                 
                 
                   VSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQI 
                 
                 
                   YKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC 
                 
                 
                   AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGV 
                 
                 
                   TQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFG 
                 
                 
                   AISSVLNDILSRLD PP EAEVQIIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECV 
                 
                 
                   LGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPRE 
                 
                 
                   GVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELD 
                 
                 
                   KYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWP 
                 
                 
                   WYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT 
                 
                 
                     
                 
                 
                   SEQ ID NO: 20 is a SARS-CoV-2 “Proline &amp; Furin Cleavage Modified” Spike 
                 
                 
                   Glycoprotein, Nucleotide Sequence 
                 
                 
                   ATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAAC 
                 
                 
                   CAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCT 
                 
                 
                   GACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTT 
                 
                 
                   TTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGG 
                 
                 
                   TTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTC 
                 
                 
                   TAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTA 
                 
                 
                   CTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAA 
                 
                 
                   TGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGA 
                 
                 
                   GTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTC 
                 
                 
                   TTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTA 
                 
                 
                   AGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCG 
                 
                 
                   TGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATT 
                 
                 
                   AACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTG 
                 
                 
                   ATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCT 
                 
                 
                   AGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGT 
                 
                 
                   GCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAA 
                 
                 
                   GGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTC 
                 
                 
                   CTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCT 
                 
                 
                   GTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTAT 
                 
                 
                   ATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAAT 
                 
                 
                   GATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCA 
                 
                 
                   GACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAG 
                 
                 
                   ATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGG 
                 
                 
                   TGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAG 
                 
                 
                   AGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAA 
                 
                 
                   GGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGG 
                 
                 
                   TTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACT 
                 
                 
                   GTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACT 
                 
                 
                   TCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTT 
                 
                 
                   CCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGAC 
                 
                 
                   ACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCA 
                 
                 
                   GGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAG 
                 
                 
                   TCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGT 
                 
                 
                   TCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACT 
                 
                 
                   CATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGAC 
                 
                 
                   TAATTCTCCTGGTAGTGCAAGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATG 
                 
                 
                   TCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAA 
                 
                 
                   ATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGT 
                 
                 
                   AGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAA 
                 
                 
                   TATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAG 
                 
                 
                   ACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAA 
                 
                 
                   TTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAG 
                 
                 
                   CAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGC 
                 
                 
                   TTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTG 
                 
                 
                   CACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGC 
                 
                 
                   TCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCA 
                 
                 
                   GGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTG 
                 
                 
                   GAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATA 
                 
                 
                   GTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACT 
                 
                 
                   TCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAG 
                 
                 
                   CTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACCCTC 
                 
                 
                   CTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGA 
                 
                 
                   CATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGC 
                 
                 
                   TGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGA 
                 
                 
                   AAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCA 
                 
                 
                   TGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCAT 
                 
                 
                   GATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGT 
                 
                 
                   TTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGT 
                 
                 
                   GTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTG 
                 
                 
                   CAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACA 
                 
                 
                   TCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTC 
                 
                 
                   AAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCG 
                 
                 
                   ATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATGGCCATGGTACATTTGGC 
                 
                 
                   TAGGTTTTATAGCTGGCTTGATTGCCATAGTAATGGTGACAATTATGCTTTGCTGTAT 
                 
                 
                   GACCAGTTGCTGTAGTTGTCTCAAGGGCTGTTGTTCTTGTGGATCCTGCTGCAAATTT 
                 
                 
                   GATGAAGACGACTCTGAGCCAGTGCTCAAAGGAGTCAAATTACATTACACATAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 21 is a SARS-Cov-2 “Proline &amp; Furin Cleavage Modified” Spike 
                 
                 
                   Glycoprotein, Nucleotide Sequence, Codon Optimized for Expression in 
                 
                 
                   Human Cells 
                 
                 
                   ATGTTCGTGTTTCTGGTGCTGCTGCCTCTGGTGAGCTCCCAGTGCGTGAACCTGACCA 
                 
                 
                   CAAGGACCCAGCTCCCCCCTGCCTATACCAATTCCTTCACAAGGGGCGTGTACTATCC 
                 
                 
                   AGACAAGGTGTTTCGCTCTAGCGTGCTGCACAGCACACAGGATCTGTTTCTGCCCTTC 
                 
                 
                   TTTTCCAACGTGACCTGGTTCCACGCCATCCATGTGAGCGGCACCAATGGCACAAAG 
                 
                 
                   AGGTTCGACAATCCTGTGCTGCCCTTCAACGATGGCGTGTACTTCGCCTCTACCGAGA 
                 
                 
                   AGAGCAACATCATCCGCGGCTGGATCTTTGGCACCACACTGGACTCCAAGACACAGT 
                 
                 
                   CTCTGCTGATCGTGAACAATGCCACCAACGTGGTCATCAAGGTGTGCGAGTTCCAGTT 
                 
                 
                   TTGTAATGATCCTTTCCTGGGCGTGTACTATCACAAGAACAATAAGAGCTGGATGGA 
                 
                 
                   GTCCGAGTTTCGCGTGTATTCCTCTGCCAACAATTGCACATTTGAGTACGTGTCCCAG 
                 
                 
                   CCATTCCTGATGGACCTGGAGGGCAAGCAGGGCAATTTCAAGAACCTGCGGGAGTTC 
                 
                 
                   GTGTTTAAGAATATCGATGGCTACTTCAAGATCTACAGCAAGCACACCCCTATCAACC 
                 
                 
                   TGGTGAGAGACCTGCCACAGGGCTTCTCCGCCCTGGAGCCTCTGGTGGATCTGCCAAT 
                 
                 
                   CGGCATCAACATCACCAGGTTTCAGACACTGCTGGCCCTGCACCGCAGCTACCTGAC 
                 
                 
                   ACCTGGCGACAGCTCCTCTGGATGGACCGCCGGGGCCGCCGCCTACTATGTGGGCTA 
                 
                 
                   TCTGCAGCCACGGACCTTCCTGCTGAAGTACAACGAGAATGGCACCATCACAGACGC 
                 
                 
                   AGTGGATTGCGCCCTGGACCCCCTGTCCGAGACCAAGTGTACACTGAAGTCTTTTACC 
                 
                 
                   GTGGAGAAGGGCATCTATCAGACATCTAATTTCCGGGTGCAGCCCACCGAGAGCATC 
                 
                 
                   GTGAGATTTCCAAATATCACAAACCTGTGCCCCTTTGGCGAGGTGTTCAACGCCACCA 
                 
                 
                   GATTCGCCAGCGTGTACGCCTGGAATCGGAAGAGAATCAGCAACTGCGTGGCCGACT 
                 
                 
                   ATTCCGTGCTGTACAACTCTGCCAGCTTCTCCACCTTTAAGTGCTATGGCGTGTCTCCC 
                 
                 
                   ACAAAGCTGAATGACCTGTGCTTTACCAACGTGTACGCCGATAGCTTCGTGATCAGG 
                 
                 
                   GGCGACGAGGTGAGACAGATCGCACCAGGCCAGACAGGCAAGATCGCCGACTACAA 
                 
                 
                   TTATAAGCTGCCCGACGATTTCACCGGCTGCGTGATCGCCTGGAACAGCAACAATCT 
                 
                 
                   GGATTCCAAAGTGGGCGGCAACTACAATTATCTGTACAGGCTGTTTCGCAAGTCCAA 
                 
                 
                   TCTGAAGCCTTTCGAGCGGGACATCAGCACAGAGATCTACCAGGCCGGCTCCACCCC 
                 
                 
                   ATGCAATGGCGTGGAGGGCTTTAACTGTTATTTCCCCCTGCAGTCTTACGGCTTCCAG 
                 
                 
                   CCTACAAACGGCGTGGGCTATCAGCCATACAGAGTGGTGGTGCTGTCCTTTGAGCTG 
                 
                 
                   CTGCACGCACCAGCAACAGTGTGCGGACCTAAGAAGTCTACCAATCTGGTGAAGAAC 
                 
                 
                   AAGTGCGTGAACTTCAACTTCAACGGCCTGACCGGCACAGGCGTGCTGACCGAGTCC 
                 
                 
                   AACAAGAAGTTCCTGCCCTTTCAGCAGTTCGGCAGAGACATCGCCGATACCACAGAC 
                 
                 
                   GCCGTGAGAGACCCCCAGACCCTGGAGATCCTGGACATCACACCTTGCTCTTTCGGC 
                 
                 
                   GGCGTGAGCGTGATCACACCTGGCACCAATACAAGCAACCAGGTGGCCGTGCTGTAT 
                 
                 
                   CAGGACGTGAATTGTACCGAGGTGCCAGTGGCCATCCACGCCGATCAGCTCACCCCC 
                 
                 
                   ACATGGAGGGTGTACTCCACCGGCTCTAACGTGTTCCAGACACGCGCCGGATGCCTG 
                 
                 
                   ATCGGAGCCGAGCATGTGAACAATTCTTATGAGTGCGACATCCCCATCGGAGCCGGC 
                 
                 
                   ATCTGTGCCAGCTACCAGACCCAGACAAACAGCCCTGGCTCCGCCAGCTCCGTGGCC 
                 
                 
                   TCTCAGAGCATCATCGCCTATACCATGAGCCTGGGGGCCGAGAATAGCGTGGCCTAC 
                 
                 
                   TCTAACAATAGCATCGCCATCCCCACCAACTTCACAATCTCCGTGACCACAGAGATCC 
                 
                 
                   TGCCCGTGAGCATGACCAAGACATCTGTGGACTGCACAATGTATATCTGTGGCGATTC 
                 
                 
                   TACCGAGTGCAGCAACCTGCTGCTGCAGTACGGCAGCTTTTGTACCCAGCTCAACCG 
                 
                 
                   GGCCCTGACAGGAATCGCAGTGGAGCAGGACAAGAACACACAGGAGGTGTTCGCCC 
                 
                 
                   AGGTGAAGCAGATCTACAAGACCCCACCCATCAAGGACTTTGGCGGCTTCAACTTCA 
                 
                 
                   GCCAGATCCTGCCAGATCCCTCCAAGCCTTCTAAGAGGAGCTTTATCGAGGACCTGCT 
                 
                 
                   GTTCAACAAGGTGACCCTGGCCGATGCCGGCTTCATCAAGCAGTATGGCGATTGCCT 
                 
                 
                   GGGCGACATCGCAGCCCGCGACCTGATCTGTGCCCAGAAGTTTAATGGCCTGACCGT 
                 
                 
                   GCTGCCTCCACTGCTGACAGATGAGATGATCGCACAGTACACATCCGCCCTGCTGGC 
                 
                 
                   CGGCACCATCACATCTGGATGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCATT 
                 
                 
                   TGCCATGCAGATGGCCTATAGATTCAACGGCATCGGCGTGACCCAGAATGTGCTGTA 
                 
                 
                   CGAGAACCAGAAGCTGATCGCCAATCAGTTTAACTCCGCCATCGGCAAGATCCAGGA 
                 
                 
                   CTCCCTGTCTAGCACAGCCTCTGCCCTGGGCAAGCTGCAGGATGTGGTGAATCAGAA 
                 
                 
                   CGCCCAGGCCCTGAATACCCTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAG 
                 
                 
                   CAGCGTGCTGAACGACATCCTGAGCCGGCTGGACCCCCCTGAGGCAGAGGTGCAGAT 
                 
                 
                   CGACAGGCTGATCACAGGCCGCCTGCAGAGCCTGCAGACCTACGTGACACAGCAGCT 
                 
                 
                   CATCAGGGCCGCCGAGATCAGAGCCTCCGCCAATCTGGCCGCCACCAAGATGTCTGA 
                 
                 
                   GTGCGTGCTGGGCCAGAGCAAGCGCGTGGACTTTTGTGGCAAGGGCTATCACCTGAT 
                 
                 
                   GTCCTTCCCACAGTCTGCCCCCCACGGAGTGGTGTTTCTGCATGTGACCTACGTGCCT 
                 
                 
                   GCCCAGGAGAAGAACTTCACCACAGCCCCAGCCATCTGCCACGATGGCAAGGCACAC 
                 
                 
                   TTTCCCCGGGAGGGCGTGTTCGTGTCTAACGGCACCCACTGGTTTGTGACACAGAGA 
                 
                 
                   AATTTCTACGAGCCTCAGATCATCACCACAGACAATACATTCGTGAGCGGCAACTGT 
                 
                 
                   GACGTGGTCATCGGCATCGTGAACAATACCGTGTATGATCCCCTGCAGCCTGAGCTG 
                 
                 
                   GACTCCTTTAAGGAGGAGCTGGATAAGTACTTCAAGAATCACACCTCTCCCGACGTG 
                 
                 
                   GATCTGGGCGACATCAGCGGCATCAATGCCTCCGTGGTGAACATCCAGAAGGAGATC 
                 
                 
                   GACAGGCTGAACGAGGTGGCCAAGAATCTGAACGAGTCCCTGATCGATCTGCAGGAG 
                 
                 
                   CTGGGCAAGTATGAGCAGTACATCAAGTGGCCATGGTATATCTGGCTGGGCTTCATC 
                 
                 
                   GCCGGCCTGATCGCCATCGTGATGGTGACCATCATGCTGTGCTGTATGACATCCTGCT 
                 
                 
                   GTTCTTGCCTGAAGGGCTGCTGTAGCTGTGGCTCCTGCTGTAAGTTTGATGAGGACGA 
                 
                 
                   TAGCGAGCCCGTGCTGAAGGGCGTGAAGCTGCACTACACCTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 22 is a SARS-CoV-2 “Proline &amp; Furin Cleavage Modified &amp; 
                 
                 
                   VSV-G TMCyt Swap” Spike Glycoprotein, Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLAL 
                 
                 
                   HRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTL 
                 
                 
                   KSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVAD 
                 
                 
                   YSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYK 
                 
                 
                   LPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVE 
                 
                 
                   GFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNF 
                 
                 
                   NGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTS 
                 
                 
                   NQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDI 
                 
                 
                   PIGAGICASYQTQTNSP GS A S SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILP 
                 
                 
                   VSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQI 
                 
                 
                   YKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC 
                 
                 
                   AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGV 
                 
                 
                   TQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFG 
                 
                 
                   AISSVLNDILSRLD PP EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECV 
                 
                 
                   LGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPRE 
                 
                 
                   GVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELD 
                 
                 
                   KYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKFFFI 
                 
                 
                   IGLIIGLFLVERVGIHECIKLKEITKKRQIYTDIEMNREGK 
                 
                 
                     
                 
                 
                   SEQ ID NO: 23 is a SARS CoV-2 “Proline &amp; Furin Cleavage Modified &amp; 
                 
                 
                   VSV-G TMCyt SWAP” Spike Glycoprotein, Nucleotide Sequence 
                 
                 
                   ATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAAC 
                 
                 
                   CAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCT 
                 
                 
                   GACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTT 
                 
                 
                   TTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGG 
                 
                 
                   TTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTC 
                 
                 
                   TAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTA 
                 
                 
                   CTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAA 
                 
                 
                   TGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGA 
                 
                 
                   GTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTC 
                 
                 
                   TTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTA 
                 
                 
                   AGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCG 
                 
                 
                   TGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATT 
                 
                 
                   AACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTG 
                 
                 
                   ATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCT 
                 
                 
                   AGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGT 
                 
                 
                   GCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAA 
                 
                 
                   GGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTC 
                 
                 
                   CTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCT 
                 
                 
                   GTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTAT 
                 
                 
                   ATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAAT 
                 
                 
                   GATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCA 
                 
                 
                   GACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAG 
                 
                 
                   ATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGG 
                 
                 
                   TGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAG 
                 
                 
                   AGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAA 
                 
                 
                   GGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGG 
                 
                 
                   TTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACT 
                 
                 
                   GTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACT 
                 
                 
                   TCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTT 
                 
                 
                   CCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGAC 
                 
                 
                   ACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCA 
                 
                 
                   GGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAG 
                 
                 
                   TCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGT 
                 
                 
                   TCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACT 
                 
                 
                   CATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGAC 
                 
                 
                   TAATTCTCCTGGTAGTGCAAGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATG 
                 
                 
                   TCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAA 
                 
                 
                   ATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGT 
                 
                 
                   AGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAA 
                 
                 
                   TATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAG 
                 
                 
                   ACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAA 
                 
                 
                   TTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAG 
                 
                 
                   CAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGC 
                 
                 
                   TTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTG 
                 
                 
                   CACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGC 
                 
                 
                   TCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCA 
                 
                 
                   GGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTG 
                 
                 
                   GAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATA 
                 
                 
                   GTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACT 
                 
                 
                   TCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAG 
                 
                 
                   CTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACCCTC 
                 
                 
                   CTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGA 
                 
                 
                   CATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGC 
                 
                 
                   TGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGA 
                 
                 
                   AAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCA 
                 
                 
                   TGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCAT 
                 
                 
                   GATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGT 
                 
                 
                   TTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGT 
                 
                 
                   GTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTG 
                 
                 
                   CAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACA 
                 
                 
                   TCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTC 
                 
                 
                   AAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCG 
                 
                 
                   ATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATTTTTCTTTATCATAGGGTT 
                 
                 
                   AATCATTGGACTATTCTTGGTTCTCCGAGTTGGTATCCATCTTTGCATTAAATTAAAGC 
                 
                 
                   ACACCAAGAAAAGACAGATTTATACAGACATAGAGATGAACCGACTTGGAAAGTAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 24 is a SARS CoV-2 “Proline &amp; Furin Cleavage Modified &amp; 
                 
                 
                   VSV-G TMCyt SWAP” Spike Glycoprotein, Nucleotide Sequence, Codon 
                 
                 
                   Optimized For Expression In Human Cells 
                 
                 
                   ATGTTCGTGTTCCTGGTGCTGCTGCCTCTGGTGAGCTCCCAGTGCGTGAACCTGACCA 
                 
                 
                   CAAGGACCCAGCTCCCCCCTGCCTATACCAATTCCTTTACAAGGGGCGTGTACTATCC 
                 
                 
                   AGACAAGGTGTTCCGCTCTAGCGTGCTGCACTCTACACAGGATCTGTTCCTGCCCTTC 
                 
                 
                   TTTAGCAACGTGACCTGGTTTCACGCCATCCATGTGAGCGGCACCAATGGCACAAAG 
                 
                 
                   CGGTTTGACAATCCTGTGCTGCCATTCAACGATGGCGTGTACTTTGCCTCCACCGAGA 
                 
                 
                   AGTCTAACATCATCAGAGGCTGGATCTTCGGCACCACACTGGACAGCAAGACACAGT 
                 
                 
                   CCCTGCTGATCGTGAACAATGCCACCAACGTGGTCATCAAGGTGTGCGAGTTTCAGTT 
                 
                 
                   CTGTAATGATCCTTTTCTGGGCGTGTACTATCACAAGAACAATAAGTCTTGGATGGAG 
                 
                 
                   AGCGAGTTCCGCGTGTATTCCTCTGCCAACAATTGTACATTCGAGTACGTGTCCCAGC 
                 
                 
                   CATTTCTGATGGACCTGGAGGGCAAGCAGGGCAACTTCAAGAACCTGCGGGAGTTCG 
                 
                 
                   TGTTCAAGAATATCGATGGCTATTTCAAGATCTACTCTAAGCACACCCCTATCAACCT 
                 
                 
                   GGTGCGCGACCTGCCACAGGGCTTTAGCGCCCTGGAGCCTCTGGTGGATCTGCCAAT 
                 
                 
                   CGGCATCAACATCACCAGGTTCCAGACACTGCTGGCCCTGCACCGCAGCTACCTGAC 
                 
                 
                   ACCTGGCGACAGCTCCTCTGGATGGACCGCCGGGGCCGCCGCCTACTATGTGGGCTA 
                 
                 
                   TCTGCAGCCACGGACCTTTCTGCTGAAGTACAACGAGAATGGCACCATCACAGACGC 
                 
                 
                   AGTGGATTGCGCCCTGGACCCCCTGAGCGAGACCAAGTGTACACTGAAGTCCTTCAC 
                 
                 
                   CGTGGAGAAGGGCATCTATCAGACATCCAATTTTCGGGTGCAGCCCACCGAGTCTAT 
                 
                 
                   CGTGAGATTCCCAAATATCACAAACCTGTGCCCCTTCGGCGAGGTGTTTAACGCCACC 
                 
                 
                   AGATTCGCCAGCGTGTACGCCTGGAATCGGAAGAGAATCTCTAACTGCGTGGCCGAC 
                 
                 
                   TATAGCGTGCTGTACAACTCTGCCAGCTTTTCCACCTTCAAGTGCTATGGCGTGTCCC 
                 
                 
                   CCACAAAGCTGAATGACCTGTGCTTCACCAACGTGTACGCCGATTCTTTTGTGATCAG 
                 
                 
                   GGGCGACGAGGTGAGACAGATCGCACCAGGCCAGACAGGCAAGATCGCCGACTACA 
                 
                 
                   ATTATAAGCTGCCCGACGATTTCACCGGCTGCGTGATCGCCTGGAACTCTAACAATCT 
                 
                 
                   GGATAGCAAAGTGGGCGGCAACTACAATTATCTGTACAGGCTGTTCCGCAAGAGCAA 
                 
                 
                   TCTGAAGCCTTTTGAGCGGGACATCTCTACAGAGATCTACCAGGCCGGCAGCACCCC 
                 
                 
                   ATGCAATGGCGTGGAGGGCTTCAACTGTTATTTTCCCCTGCAGTCCTACGGCTTTCAG 
                 
                 
                   CCTACCAACGGCGTGGGCTATCAGCCATACAGAGTGGTGGTGCTGAGCTTCGAGCTG 
                 
                 
                   CTGCACGCACCAGCAACAGTGTGCGGACCTAAGAAGTCCACCAATCTGGTGAAGAAC 
                 
                 
                   AAGTGCGTGAACTTCAACTTCAACGGCCTGACCGGCACAGGCGTGCTGACCGAGTCC 
                 
                 
                   AATAAGAAGTTTCTGCCCTTCCAGCAGTTTGGCCGGGACATCGCCGATACCACAGAC 
                 
                 
                   GCCGTGAGAGACCCCCAGACCCTGGAGATCCTGGACATCACACCTTGCTCCTTCGGC 
                 
                 
                   GGCGTGTCTGTGATCACACCTGGCACCAATACAAGCAACCAGGTGGCCGTGCTGTAT 
                 
                 
                   CAGGACGTGAATTGTACCGAGGTGCCAGTGGCCATCCACGCCGATCAGCTCACCCCC 
                 
                 
                   ACATGGCGGGTGTACTCCACCGGCTCTAACGTGTTCCAGACAAGAGCCGGCTGCCTG 
                 
                 
                   ATCGGAGCCGAGCATGTGAACAATTCCTATGAGTGCGACATCCCCATCGGAGCCGGC 
                 
                 
                   ATCTGTGCCTCTTACCAGACCCAGACAAACAGCCCTGGCTCCGCCAGCTCCGTGGCCT 
                 
                 
                   CTCAGAGCATCATCGCCTATACCATGAGCCTGGGGGCCGAGAACAGCGTGGCCTACT 
                 
                 
                   CTAACAATAGCATCGCCATCCCCACCAACTTTACAATCTCTGTGACCACAGAGATCCT 
                 
                 
                   GCCTGTGAGCATGACCAAGACATCCGTGGACTGCACAATGTATATCTGTGGCGATTC 
                 
                 
                   CACCGAGTGCTCTAACCTGCTGCTGCAGTACGGCAGCTTCTGTACCCAGCTCAACCGG 
                 
                 
                   GCCCTGACAGGAATCGCAGTGGAGCAGGACAAGAACACACAGGAGGTGTTTGCCCA 
                 
                 
                   GGTGAAGCAGATCTACAAGACCCCACCCATCAAGGACTTCGGCGGCTTTAATTTCTCC 
                 
                 
                   CAGATCCTGCCAGATCCCTCCAAGCCATCTAAGCGGAGCTTCATCGAGGACCTGCTGT 
                 
                 
                   TTAACAAGGTGACCCTGGCCGATGCCGGCTTTATCAAGCAGTATGGCGATTGCCTGG 
                 
                 
                   GCGACATCGCCGCCAGAGACCTGATCTGTGCCCAGAAGTTCAATGGCCTGACCGTGC 
                 
                 
                   TGCCTCCACTGCTGACAGATGAGATGATCGCACAGTACACAAGCGCCCTGCTGGCCG 
                 
                 
                   GCACCATCACATCCGGATGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCCTTCG 
                 
                 
                   CCATGCAGATGGCCTATAGGTTTAACGGCATCGGCGTGACCCAGAATGTGCTGTACG 
                 
                 
                   AGAACCAGAAGCTGATCGCCAATCAGTTCAACTCCGCCATCGGCAAGATCCAGGACA 
                 
                 
                   GCCTGTCTAGCACAGCCTCCGCCCTGGGCAAGCTGCAGGATGTGGTGAATCAGAACG 
                 
                 
                   CCCAGGCCCTGAATACCCTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAGCA 
                 
                 
                   GCGTGCTGAACGACATCCTGAGCCGGCTGGACCCCCCTGAGGCAGAGGTGCAGATCG 
                 
                 
                   ACAGGCTGATCACAGGCCGCCTGCAGTCTCTGCAGACCTATGTGACACAGCAGCTCA 
                 
                 
                   TCAGGGCCGCCGAGATCAGAGCCAGCGCCAATCTGGCCGCCACCAAGATGTCCGAGT 
                 
                 
                   GCGTGCTGGGCCAGTCTAAGCGCGTGGACTTCTGTGGCAAGGGCTATCACCTGATGA 
                 
                 
                   GCTTTCCACAGTCCGCCCCCCACGGAGTGGTGTTCCTGCATGTGACCTACGTGCCTGC 
                 
                 
                   CCAGGAGAAGAACTTTACCACAGCCCCAGCCATCTGCCACGATGGCAAGGCACACTT 
                 
                 
                   CCCCAGGGAGGGCGTGTTCGTGAGCAACGGCACCCACTGGTTCGTGACACAGCGCAA 
                 
                 
                   CTTCTACGAGCCTCAGATCATCACCACAGACAATACATTCGTGTCTGGCAACTGTGAC 
                 
                 
                   GTGGTCATCGGCATCGTGAACAATACCGTGTATGATCCCCTGCAGCCTGAGCTGGAC 
                 
                 
                   AGCTTCAAGGAGGAGCTGGATAAGTACTTTAAGAATCACACCTCCCCCGACGTGGAT 
                 
                 
                   CTGGGCGACATCTCTGGCATCAATGCCAGCGTGGTGAACATCCAGAAGGAGATCGAC 
                 
                 
                   AGGCTGAACGAGGTGGCCAAGAATCTGAACGAGAGCCTGATCGATCTGCAGGAGCT 
                 
                 
                   GGGCAAGTATGAGCAGTACATCAAGTTCTTTTTCATCATCGGCCTGATCATCGGCCTG 
                 
                 
                   TTCCTGGTGCTGCGCGTGGGCATCCACCTGTGCATCAAGCTGAAGCACACCAAGAAG 
                 
                 
                   AGGCAGATCTACACAGACATCGAGATGAACCGCCTGGGCAAGTGA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 25 is a SARS-CoV-2 Spike Glycoprotein, with VSV-G TMCyt 
                 
                 
                   SWAP Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLAL 
                 
                 
                   HRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTL 
                 
                 
                   KSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVAD 
                 
                 
                   YSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYK 
                 
                 
                   LPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVE 
                 
                 
                   GFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNF 
                 
                 
                   NGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTS 
                 
                 
                   NQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDI 
                 
                 
                   PIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILP 
                 
                 
                   VSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQI 
                 
                 
                   YKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLIC 
                 
                 
                   AQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGV 
                 
                 
                   TQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFG 
                 
                 
                   AISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSEC 
                 
                 
                   VLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPR 
                 
                 
                   EGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEEL 
                 
                 
                   DKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIK FF   
                 
                 
                   
                     FIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK 
                   
                 
                 
                     
                 
                 
                   SEQ ID NO: 26 is a SARS-CoV-2 Spike Glycoprotein with VSV-G TMCyt 
                 
                 
                   SWAP, Nucleotide Sequence 
                 
                 
                   CTCGAGGTTTAAACGAATTCCGCCACCATGTTTGTTTTTCTTGTTTTATTGCCACTAGT 
                 
                 
                   CTCTAGTCAGTGTGTTAATCTTACAACCAGAACTCAATTACCCCCTGCATACACTAAT 
                 
                 
                   TCTTTCACACGTGGTGTTTATTACCCTGACAAAGTTTTCAGATCCTCAGTTTTACATTC 
                 
                 
                   AACTCAGGACTTGTTCTTACCTTTCTTTTCCAATGTTACTTGGTTCCATGCTATACATG 
                 
                 
                   TCTCTGGGACCAATGGTACTAAGAGGTTTGATAACCCTGTCCTACCATTTAATGATGG 
                 
                 
                   TGTTTATTTTGCTTCCACTGAGAAGTCTAACATAATAAGAGGCTGGATTTTTGGTACT 
                 
                 
                   ACTTTAGATTCGAAGACCCAGTCCCTACTTATTGTTAATAACGCTACTAATGTTGTTA 
                 
                 
                   TTAAAGTCTGTGAATTTCAATTTTGTAATGATCCATTTTTGGGTGTTTATTACCACAAA 
                 
                 
                   AACAACAAAAGTTGGATGGAAAGTGAGTTCAGAGTTTATTCTAGTGCGAATAATTGC 
                 
                 
                   ACTTTTGAATATGTCTCTCAGCCTTTTCTTATGGACCTTGAAGGAAAACAGGGTAATT 
                 
                 
                   TCAAAAATCTTAGGGAATTTGTGTTTAAGAATATTGATGGTTATTTTAAAATATATTC 
                 
                 
                   TAAGCACACGCCTATTAATTTAGTGCGTGATCTCCCTCAGGGTTTTTCGGCTTTAGAA 
                 
                 
                   CCATTGGTAGATTTGCCAATAGGTATTAACATCACTAGGTTTCAAACTTTACTTGCTTT 
                 
                 
                   ACATAGAAGTTATTTGACTCCTGGTGATTCTTCTTCAGGTTGGACAGCTGGTGCTGCA 
                 
                 
                   GCTTATTATGTGGGTTATCTTCAACCTAGGACTTTTCTATTAAAATATAATGAAAATG 
                 
                 
                   GAACCATTACAGATGCTGTAGACTGTGCACTTGACCCTCTCTCAGAAACAAAGTGTA 
                 
                 
                   CGTTGAAATCCTTCACTGTAGAAAAAGGAATCTATCAAACTTCTAACTTTAGAGTCCA 
                 
                 
                   ACCAACAGAATCTATTGTTAGATTTCCTAATATTACAAACTTGTGCCCTTTTGGTGAA 
                 
                 
                   GTTTTTAACGCCACCAGATTTGCATCTGTTTATGCTTGGAACAGGAAGAGAATCAGCA 
                 
                 
                   ACTGTGTTGCTGATTATTCTGTCCTATATAATTCCGCATCATTTTCCACTTTTAAGTGT 
                 
                 
                   TATGGAGTGTCTCCTACTAAATTAAATGATCTCTGCTTTACTAATGTCTATGCAGATTC 
                 
                 
                   ATTTGTAATTAGAGGTGATGAAGTCAGACAAATCGCTCCAGGGCAAACTGGAAAGAT 
                 
                 
                   TGCTGATTATAATTATAAATTACCAGATGATTTTACAGGCTGCGTTATAGCTTGGAAT 
                 
                 
                   TCTAACAATCTTGATTCTAAGGTTGGTGGTAATTATAATTACCTGTATAGATTGTTTA 
                 
                 
                   GGAAGTCTAATCTCAAACCTTTTGAGAGAGATATTTCAACTGAAATCTATCAGGCCG 
                 
                 
                   GTAGCACACCTTGTAATGGTGTTGAAGGTTTTAATTGTTACTTTCCTTTACAATCATAT 
                 
                 
                   GGTTTCCAACCCACTAATGGTGTTGGTTACCAACCATACAGAGTAGTAGTACTTTCTT 
                 
                 
                   TTGAACTTCTACATGCACCAGCAACTGTTTGTGGACCTAAAAAGTCTACTAATTTGGT 
                 
                 
                   TAAAAACAAATGTGTCAATTTCAACTTCAATGGTTTAACAGGCACAGGTGTTCTTACT 
                 
                 
                   GAGTCTAACAAAAAGTTTCTGCCTTTCCAACAATTTGGCAGAGACATTGCTGACACTA 
                 
                 
                   CTGATGCTGTCCGTGATCCACAGACACTTGAGATTCTTGACATTACACCATGTTCTTTT 
                 
                 
                   GGTGGTGTCAGTGTTATAACACCAGGAACAAATACTTCTAACCAGGTTGCTGTTCTTT 
                 
                 
                   ATCAGGATGTTAACTGCACAGAAGTCCCTGTTGCTATTCATGCAGATCAACTTACTCC 
                 
                 
                   TACTTGGCGTGTTTATTCTACAGGTTCTAATGTTTTTCAAACACGTGCAGGCTGTTTAA 
                 
                 
                   TAGGGGCTGAACATGTCAACAACTCATATGAGTGTGACATACCCATTGGTGCAGGTA 
                 
                 
                   TATGCGCTAGTTATCAGACTCAGACTAATTCTCCTCGGCGGGCACGTAGTGTAGCTAG 
                 
                 
                   TCAATCCATCATTGCCTACACTATGTCACTTGGTGCAGAAAATTCAGTTGCTTACTCT 
                 
                 
                   AATAACTCTATTGCCATACCCACAAATTTTACTATTAGTGTTACCACAGAAATTCTAC 
                 
                 
                   CAGTGTCTATGACCAAGACATCAGTAGATTGTACAATGTACATTTGTGGTGATTCAAC 
                 
                 
                   TGAATGCAGCAATCTTTTGTTGCAATATGGCAGTTTTTGTACACAATTAAACCGTGCT 
                 
                 
                   TTAACTGGAATAGCTGTTGAACAAGACAAAAACACCCAAGAAGTTTTTGCACAAGTC 
                 
                 
                   AAACAAATTTACAAAACACCACCAATTAAAGATTTTGGTGGTTTTAATTTTTCACAAA 
                 
                 
                   TATTACCAGATCCATCAAAACCAAGCAAGAGGTCATTTATTGAAGATCTACTTTTCAA 
                 
                 
                   CAAAGTGACACTTGCAGATGCTGGCTTCATCAAACAATATGGTGATTGCCTTGGTGAT 
                 
                 
                   ATTGCTGCTAGAGACCTCATTTGTGCACAAAAGTTTAACGGCCTTACTGTTTTGCCAC 
                 
                 
                   CTTTGCTCACAGATGAAATGATTGCTCAATACACTTCTGCACTGTTAGCGGGTACAAT 
                 
                 
                   CACTTCTGGTTGGACCTTTGGTGCAGGTGCTGCATTACAAATACCATTTGCTATGCAA 
                 
                 
                   ATGGCTTATAGGTTTAATGGTATTGGAGTTACACAGAATGTTCTCTATGAGAACCAAA 
                 
                 
                   AATTGATTGCCAACCAATTTAATAGTGCTATTGGCAAAATTCAAGACTCACTTTCTTC 
                 
                 
                   CACAGCAAGTGCACTTGGAAAACTTCAAGATGTGGTCAACCAAAATGCACAAGCTTT 
                 
                 
                   AAACACGCTTGTTAAACAACTTAGCTCCAATTTTGGTGCAATTTCAAGTGTTTTAAAT 
                 
                 
                   GATATCCTTTCACGTCTTGACAAAGTTGAGGCTGAAGTGCAAATTGATAGGTTGATCA 
                 
                 
                   CAGGCAGACTTCAAAGTTTGCAGACATATGTGACTCAACAATTAATTAGAGCTGCAG 
                 
                 
                   AAATCAGAGCTTCTGCTAATCTTGCTGCTACTAAAATGTCAGAGTGTGTACTTGGACA 
                 
                 
                   ATCAAAAAGAGTTGATTTTTGTGGAAAGGGCTATCATCTTATGTCCTTCCCTCAGTCA 
                 
                 
                   GCACCTCATGGTGTAGTCTTCTTGCATGTGACTTATGTCCCTGCACAAGAAAAGAACT 
                 
                 
                   TCACAACTGCTCCTGCCATTTGTCATGATGGAAAAGCACACTTTCCTCGTGAAGGTGT 
                 
                 
                   CTTTGTTTCAAATGGCACACACTGGTTTGTAACACAAAGGAATTTTTATGAACCACAA 
                 
                 
                   ATCATTACTACAGACAACACATTTGTGTCTGGTAACTGTGATGTTGTAATAGGAATTG 
                 
                 
                   TCAACAACACAGTTTATGATCCTTTGCAACCTGAATTAGACTCATTCAAGGAGGAGTT 
                 
                 
                   AGATAAATATTTTAAGAATCATACATCACCAGATGTTGATTTAGGTGACATCTCTGGC 
                 
                 
                   ATTAATGCTTCAGTTGTAAACATTCAAAAAGAAATTGACCGCCTCAATGAGGTTGCC 
                 
                 
                   AAGAATTTAAATGAATCTCTCATCGATCTCCAAGAACTTGGAAAGTATGAGCAGTAT 
                 
                 
                   ATAAAATGGCCATTTTTCTTTATCATAGGGTTAATCATTGGACTATTCTTGGTTCTCCG 
                 
                 
                   AGTTGGTATCCATCTTTGCATTAAATTAAAGCACACCAAGAAAAGACAGATTTATAC 
                 
                 
                   AGACATAGAGATGAACCGACTTGGAAAGTAAGAATTCCACGTGGGATCC 
                 
                 
                     
                 
                 
                   SEQ ID NO: 27 is a SARS CoV-2 Spike Glycoprotein with VSV-G TMCyt 
                 
                 
                   SWAP, Nucleotide Sequence, Codon Optimized For Expression In Human Cells 
                 
                 
                   CTCGAGGTTTAAACGAATTCCGCCACCATGTTCGTGTTCCTGGTGCTGCTGCCCCTGG 
                 
                 
                   TGTCCAGCCAGTGCGTGAATCTGACCACCCGGACCCAACTGCCTCCCGCCTACACAA 
                 
                 
                   ACTCTTTCACCAGAGGGGTTTATTACCCCGATAAGGTGTTCAGAAGCTCAGTGCTTCA 
                 
                 
                   TTCTACCCAGGACCTGTTTCTGCCTTTTTTCAGCAACGTCACATGGTTCCACGCCATCC 
                 
                 
                   ACGTCAGCGGAACCAACGGCACGAAGCGGTTCGACAATCCTGTGCTGCCTTTTAACG 
                 
                 
                   ACGGCGTCTACTTTGCCAGCACGGAAAAGAGCAACATTATCCGGGGATGGATCTTCG 
                 
                 
                   GCACCACCCTGGACTCTAAAACCCAGAGCCTGTTGATCGTGAACAACGCAACCAATG 
                 
                 
                   TGGTGATCAAGGTCTGCGAGTTCCAATTTTGCAACGATCCTTTCCTGGGCGTGTACTA 
                 
                 
                   CCACAAGAACAACAAGTCTTGGATGGAATCTGAGTTCCGCGTCTACAGCAGCGCAAA 
                 
                 
                   CAACTGCACATTTGAGTACGTGTCTCAGCCTTTTCTGATGGACCTGGAAGGAAAGCA 
                 
                 
                   GGGAAATTTCAAGAACCTGCGGGAGTTCGTGTTCAAGAACATCGACGGCTACTTCAA 
                 
                 
                   GATCTACAGCAAGCACACCCCCATCAACCTCGTGAGAGACCTGCCCCAGGGCTTCAG 
                 
                 
                   CGCCCTGGAACCCCTGGTGGACCTTCCCATAGGAATCAACATCACACGGTTCCAGAC 
                 
                 
                   ACTGCTGGCCCTGCATAGAAGCTACCTGACCCCTGGAGATTCTAGCAGCGGCTGGAC 
                 
                 
                   CGCCGGCGCTGCCGCTTACTACGTCGGATACCTGCAGCCTAGAACCTTCCTGTTGAAG 
                 
                 
                   TACAACGAGAACGGCACCATCACAGATGCCGTGGACTGCGCCCTGGACCCCCTGAGC 
                 
                 
                   GAGACAAAGTGCACCCTGAAGAGCTTCACCGTGGAGAAGGGCATCTACCAGACAAG 
                 
                 
                   CAACTTCAGAGTGCAGCCTACCGAGTCAATCGTGAGATTCCCAAACATCACCAACCT 
                 
                 
                   TTGTCCTTTCGGCGAGGTATTTAACGCCACCCGGTTCGCCAGCGTGTACGCCTGGAAT 
                 
                 
                   AGGAAGCGGATCAGCAACTGCGTGGCCGATTACAGCGTGCTCTATAACAGCGCCAGT 
                 
                 
                   TTTAGCACTTTCAAGTGCTACGGAGTCTCTCCTACAAAGCTGAACGACCTGTGCTTCA 
                 
                 
                   CCAACGTGTATGCCGACAGCTTCGTCATCCGGGGCGACGAGGTGCGACAGATCGCTC 
                 
                 
                   CTGGCCAGACCGGCAAGATAGCCGACTACAACTACAAGCTGCCTGACGACTTCACAG 
                 
                 
                   GCTGCGTGATCGCTTGGAACAGCAACAATCTGGATAGCAAAGTGGGCGGCAACTATA 
                 
                 
                   ACTACCTGTACAGACTGTTCCGGAAGTCCAATCTCAAGCCGTTTGAGAGAGACATCA 
                 
                 
                   GCACCGAAATCTACCAGGCTGGATCTACACCCTGCAACGGCGTCGAAGGCTTCAATT 
                 
                 
                   GTTACTTCCCTCTGCAATCTTACGGCTTCCAGCCCACCAACGGCGTGGGCTACCAGCC 
                 
                 
                   CTACAGAGTGGTTGTGCTGAGCTTCGAGCTGCTGCACGCCCCAGCTACAGTGTGCGG 
                 
                 
                   CCCTAAGAAATCTACAAACCTGGTCAAGAACAAGTGTGTGAACTTCAACTTCAATGG 
                 
                 
                   CCTGACGGGCACCGGCGTGCTGACAGAGAGCAACAAGAAGTTCCTGCCTTTCCAGCA 
                 
                 
                   ATTTGGCAGAGACATCGCCGACACCACCGACGCCGTGCGCGACCCTCAGACCCTGGA 
                 
                 
                   AATTCTGGACATCACCCCATGTTCTTTCGGCGGCGTGTCCGTCATTACGCCAGGCACC 
                 
                 
                   AATACCAGCAACCAGGTGGCCGTGCTTTATCAGGATGTGAATTGTACCGAAGTTCCT 
                 
                 
                   GTTGCAATCCACGCCGACCAACTGACCCCCACATGGAGAGTGTACTCTACCGGCAGC 
                 
                 
                   AACGTGTTCCAAACGAGAGCCGGATGCCTGATTGGAGCTGAGCATGTGAACAACAGC 
                 
                 
                   TACGAGTGCGATATTCCAATCGGAGCCGGCATCTGCGCCTCCTACCAAACACAAACC 
                 
                 
                   AACTCCCCTCGTAGAGCGAGAAGCGTGGCCTCTCAGAGCATCATCGCCTACACCATG 
                 
                 
                   AGCCTGGGTGCCGAAAACTCCGTGGCTTACTCCAACAACAGCATCGCCATCCCTACA 
                 
                 
                   AATTTCACCATCAGCGTGACAACCGAGATCCTGCCTGTGTCCATGACCAAGACCAGC 
                 
                 
                   GTGGACTGCACGATGTACATCTGCGGAGATAGCACCGAGTGCAGCAATCTGCTACTG 
                 
                 
                   CAGTATGGCAGCTTCTGCACCCAACTGAACAGAGCACTGACCGGCATTGCTGTGGAA 
                 
                 
                   CAGGACAAGAATACCCAGGAGGTGTTCGCCCAAGTGAAGCAGATTTACAAGACACCC 
                 
                 
                   CCTATCAAGGACTTCGGAGGCTTCAACTTCAGCCAGATCCTGCCTGACCCTAGCAAGC 
                 
                 
                   CAAGCAAAAGATCCTTTATCGAAGATCTGCTGTTTAACAAGGTGACACTGGCCGATG 
                 
                 
                   CCGGCTTTATCAAGCAGTACGGCGACTGCCTGGGAGACATCGCCGCCAGAGACCTGA 
                 
                 
                   TCTGTGCTCAGAAATTTAACGGGCTGACCGTGCTGCCACCTCTGCTGACAGATGAGAT 
                 
                 
                   GATCGCTCAGTACACCAGCGCCCTGCTGGCCGGCACAATTACCTCCGGCTGGACCTTC 
                 
                 
                   GGAGCCGGAGCCGCCCTGCAGATCCCCTTCGCCATGCAGATGGCCTACCGGTTCAAT 
                 
                 
                   GGCATCGGCGTCACCCAAAACGTGCTCTATGAGAACCAGAAGCTGATCGCAAACCAG 
                 
                 
                   TTCAACTCCGCCATCGGTAAGATCCAGGACAGTCTGAGCAGCACGGCGTCTGCCCTG 
                 
                 
                   GGCAAGCTCCAGGACGTGGTGAACCAGAACGCCCAGGCCCTTAACACCCTGGTGAAA 
                 
                 
                   CAACTGAGCAGCAACTTCGGTGCCATTTCCAGCGTTCTCAATGACATCCTGAGCAGAC 
                 
                 
                   TGGATAAGGTGGAAGCCGAGGTGCAGATCGACCGGCTGATCACCGGACGGCTGCAG 
                 
                 
                   AGCCTGCAGACGTACGTGACCCAGCAATTAATCAGAGCTGCCGAGATCAGAGCCAGC 
                 
                 
                   GCCAATCTGGCTGCCACCAAAATGAGCGAATGTGTGCTGGGCCAGTCAAAGAGAGTG 
                 
                 
                   GATTTTTGTGGCAAAGGCTACCACCTGATGTCCTTCCCTCAGTCTGCCCCTCACGGCG 
                 
                 
                   TGGTGTTCCTCCATGTGACCTATGTGCCTGCTCAGGAGAAGAACTTTACCACAGCCCC 
                 
                 
                   TGCTATCTGCCACGACGGAAAGGCCCACTTCCCCAGAGAGGGCGTGTTTGTGTCCAA 
                 
                 
                   TGGCACACACTGGTTCGTGACCCAAAGAAACTTCTACGAGCCCCAGATCATCACCAC 
                 
                 
                   AGACAACACCTTCGTGAGCGGCAACTGCGACGTGGTGATCGGCATCGTGAACAACAC 
                 
                 
                   AGTGTACGACCCCCTGCAACCTGAGCTGGACAGCTTCAAAGAGGAACTGGACAAATA 
                 
                 
                   CTTCAAGAATCACACCAGCCCTGATGTGGATCTGGGCGACATCAGCGGCATCAACGC 
                 
                 
                   CAGCGTCGTGAACATCCAGAAGGAAATCGACAGACTGAACGAAGTGGCCAAGAACC 
                 
                 
                   TGAACGAGAGCCTCATCGATCTGCAGGAGCTGGGCAAGTACGAGCAGTACATCAAAT 
                 
                 
                   GGCCTTTCTTCTTCATCATCGGCCTGATTATCGGCCTGTTCCTCGTGCTGAGAGTGGGC 
                 
                 
                   ATCCACCTGTGCATCAAGCTTAAGCACACAAAAAAGCGGCAGATTTACACCGACATC 
                 
                 
                   GAGATGAACCGGCTGGGCAAATGAGAATTCCACGTGGGATCC 
                 
                 
                     
                 
                 
                   SEQ ID NO: 28 is a SARS-CoV-2 Beta Variant Spike Glycoprotein, with 
                 
                 
                   VSV-G TMCyt SWAP Amino Acid Sequence 
                 
                 
                   MFVFLVLLPLVSSQCVNFTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSN 
                 
                 
                   VTWFHAIHVSGTNGTKRFANPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNN 
                 
                 
                   ATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEG 
                 
                 
                   KQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRGLPQGFSALEPLVDLPIGINITRFQTLHRS 
                 
                 
                   YLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFT 
                 
                 
                   VEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVL 
                 
                 
                   YNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGNIADYNYKLPDD 
                 
                 
                   FTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNC 
                 
                 
                   YFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLT 
                 
                 
                   GTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVA 
                 
                 
                   VLYQGVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAG 
                 
                 
                   ICASYQTQTNSPGSASSVASQSIIAYTMSLGVENSVAYSNNSIAIPTNFTISVTTEILPVSMT 
                 
                 
                   KTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTP 
                 
                 
                   PIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQK 
                 
                 
                   FNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQN 
                 
                 
                   VLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISS 
                 
                 
                   VLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQ 
                 
                 
                   SKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVF 
                 
                 
                   VSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYF 
                 
                 
                   KNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKFFFIIGLI 
                 
                 
                   IGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK 
                 
                 
                     
                 
                 
                   SEQ ID NO: 29 is a SARS-CoV-2 Beta Variant Spike Glycoprotein with VSV- 
                 
                 
                   G TMCyt SWAP, Nucleotide Sequence 
                 
                 
                   ATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATTTTACAACC 
                 
                 
                   AGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCTG 
                 
                 
                   ACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTTT 
                 
                 
                   TCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGGT 
                 
                 
                   TTGCTAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTCT 
                 
                 
                   AACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTAC 
                 
                 
                   TTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAAT 
                 
                 
                   GATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGAG 
                 
                 
                   TTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTCT 
                 
                 
                   TATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTAA 
                 
                 
                   GAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCGT 
                 
                 
                   GGTCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATTA 
                 
                 
                   ACATCACTAGGTTTCAAACTTTACATAGAAGTTATTTGACTCCTGGTGATTCTTCTTCA 
                 
                 
                   GGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCTAGGACTTTTC 
                 
                 
                   TATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGTGCACTTGACC 
                 
                 
                   CTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAAGGAATCTATC 
                 
                 
                   AAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTCCTAATATTAC 
                 
                 
                   AAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCTGTTTATGCTT 
                 
                 
                   GGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTATATAATTCCGC 
                 
                 
                   ATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAATGATCTCTGCT 
                 
                 
                   TTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCAGACAAATCGC 
                 
                 
                   TCCAGGGCAAACTGGAAATATTGCTGATTATAATTATAAATTACCAGATGATTTTACA 
                 
                 
                   GGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGGTGGTAATTATA 
                 
                 
                   ATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAGAGAGATATTTC 
                 
                 
                   AACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTAAAGGTTTTAATTGT 
                 
                 
                   TACTTTCCTTTACAATCATATGGTTTCCAACCCACTTATGGTGTTGGTTACCAACCATA 
                 
                 
                   CAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACTGTTTGTGGACCT 
                 
                 
                   AAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACTTCAATGGTTTAA 
                 
                 
                   CAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTTCCAACAATTTGG 
                 
                 
                   CAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGACACTTGAGATTCTT 
                 
                 
                   GACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCAGGAACAAATACTT 
                 
                 
                   CTAACCAGGTTGCTGTTCTTTATCAGGGTGTTAACTGCACAGAAGTCCCTGTTGCTAT 
                 
                 
                   TCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGTTCTAATGTTTTTC 
                 
                 
                   AAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACTCATATGAGTGTG 
                 
                 
                   ACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGACTAATTCTCCTGG 
                 
                 
                   TAGTGCAAGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATGTCACTTGGTGTA 
                 
                 
                   GAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAAATTTTACTATTA 
                 
                 
                   GTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGTAGATTGTACAAT 
                 
                 
                   GTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAATATGGCAGTTTT 
                 
                 
                   TGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAGACAAAAACACC 
                 
                 
                   CAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAATTAAAGATTTT 
                 
                 
                   GGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAGCAAGAGGTCAT 
                 
                 
                   TTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGCTTCATCAAACA 
                 
                 
                   ATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTGCACAAAAGTTT 
                 
                 
                   AACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGCTCAATACACTT 
                 
                 
                   CTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCAGGTGCTGCATT 
                 
                 
                   ACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTGGAGTTACACAG 
                 
                 
                   AATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATAGTGCTATTGGCA 
                 
                 
                   AAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACTTCAAGATGTGGT 
                 
                 
                   CAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAGCTCCAATTTTGGT 
                 
                 
                   GCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACCCTCCTGAGGCTGAAG 
                 
                 
                   TGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGACATATGTGACTC 
                 
                 
                   AACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGCTGCTACTAAAAT 
                 
                 
                   GTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGAAAGGGCTATCA 
                 
                 
                   TCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCATGTGACTTATG 
                 
                 
                   TCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCATGATGGAAAAG 
                 
                 
                   CACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGTTTGTAACACA 
                 
                 
                   AAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGTGTCTGGTAAC 
                 
                 
                   TGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTGCAACCTGAAT 
                 
                 
                   TAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACATCACCAGATG 
                 
                 
                   TTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTCAAAAAGAAAT 
                 
                 
                   TGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCGATCTCCAAGAA 
                 
                 
                   CTTGGAAAGTATGAGCAGTATATAAAATTTTTCTTTATCATAGGGTTAATCATTGGAC 
                 
                 
                   TATTCTTGGTTCTCCGAGTTGGTATCCATCTTTGCATTAAATTAAAGCACACCAAGAA 
                 
                 
                   AAGACAGATTTATACAGACATAGAGATGAACCGACTTGGAAAGTAA 
                 
                 
                     
                 
                 
                   SEQ ID NO: 30 is a SARS CoV-2 Beta Variant Spike Glycoprotein with VSV-G 
                 
                 
                   TMCyt SWAP, Nucleotide Sequence, Codon Optimized For Expression In Human 
                 
                 
                   Cells 
                 
                 
                   ATGTTCGTGTTCCTGGTGCTGCTGCCTCTGGTGAGCTCCCAGTGCGTGAACTTCACCA 
                 
                 
                   CAAGGACCCAGCTCCCCCCTGCCTACACCAATTCCTTTACAAGGGGCGTGTACTATCC 
                 
                 
                   AGACAAGGTGTTCCGCTCTAGCGTGCTGCACTCTACACAGGATCTGTTCCTGCCCTTC 
                 
                 
                   TTTAGCAACGTGACCTGGTTTCACGCCATCCATGTGAGCGGCACCAATGGCACAAAG 
                 
                 
                   CGGTTTGCCAATCCTGTGCTGCCATTCAACGACGGCGTGTATTTTGCCTCCACCGAGA 
                 
                 
                   AGTCTAACATCATCAGAGGCTGGATCTTCGGCACCACACTGGATAGCAAGACACAGT 
                 
                 
                   CCCTGCTGATCGTGAACAATGCCACCAACGTGGTCATCAAGGTGTGCGAGTTTCAGTT 
                 
                 
                   CTGTAATGACCCTTTTCTGGGCGTGTACTATCACAAGAACAATAAGTCTTGGATGGAG 
                 
                 
                   AGCGAGTTCCGCGTGTACTCCTCTGCCAACAATTGTACATTCGAGTACGTGAGCCAGC 
                 
                 
                   CATTTCTGATGGACCTGGAGGGCAAGCAGGGCAACTTCAAGAACCTGCGGGAGTTCG 
                 
                 
                   TGTTCAAGAATATCGATGGCTATTTCAAGATCTACTCTAAGCACACCCCTATCAACCT 
                 
                 
                   GGTGCGCGGACTGCCACAGGGCTTTAGCGCCCTGGAGCCTCTGGTGGACCTGCCAAT 
                 
                 
                   CGGCATCAACATCACCAGGTTCCAGACACTGCACCGCAGCTACCTGACACCTGGCGA 
                 
                 
                   TAGCTCCTCTGGATGGACCGCCGGGGCCGCCGCCTACTATGTGGGCTACCTGCAGCC 
                 
                 
                   ACGGACCTTTCTGCTGAAGTATAACGAGAATGGCACCATCACAGACGCAGTGGATTG 
                 
                 
                   CGCCCTGGACCCCCTGAGCGAGACCAAGTGTACACTGAAGTCCTTCACCGTGGAGAA 
                 
                 
                   GGGCATCTACCAGACATCCAATTTTCGGGTGCAGCCCACCGAGTCTATCGTGAGATTC 
                 
                 
                   CCAAATATCACAAACCTGTGCCCCTTCGGCGAGGTGTTTAACGCCACCAGATTCGCCA 
                 
                 
                   GCGTGTATGCCTGGAATCGGAAGAGAATCTCTAACTGCGTGGCCGACTATAGCGTGC 
                 
                 
                   TGTACAACTCTGCCAGCTTTTCCACCTTCAAGTGCTATGGCGTGTCCCCCACAAAGCT 
                 
                 
                   GAATGACCTGTGCTTCACCAACGTGTATGCCGATTCTTTTGTGATCAGGGGCGACGAG 
                 
                 
                   GTGAGACAGATCGCACCAGGCCAGACAGGCAATATCGCCGACTACAACTATAAGCTG 
                 
                 
                   CCCGACGATTTCACCGGCTGCGTGATCGCCTGGAACAGCAACAATCTGGATAGCAAA 
                 
                 
                   GTGGGCGGCAACTACAATTATCTGTACAGGCTGTTCCGCAAGAGCAACCTGAAGCCT 
                 
                 
                   TTTGAGCGGGACATCTCTACAGAGATCTACCAGGCCGGCAGCACCCCATGCAATGGC 
                 
                 
                   GTGAAGGGCTTCAACTGTTATTTTCCCCTGCAGTCCTACGGCTTTCAGCCTACCTATG 
                 
                 
                   GCGTGGGCTATCAGCCATACAGAGTGGTGGTGCTGAGCTTCGAGCTGCTGCACGCAC 
                 
                 
                   CAGCAACAGTGTGCGGACCTAAGAAGTCCACCAATCTGGTGAAGAACAAGTGCGTGA 
                 
                 
                   ACTTCAACTTCAACGGCCTGACCGGCACAGGCGTGCTGACCGAGTCCAATAAGAAGT 
                 
                 
                   TTCTGCCCTTCCAGCAGTTTGGCCGGGACATCGCCGATACCACAGACGCCGTGAGAG 
                 
                 
                   ACCCCCAGACCCTGGAGATCCTGGACATCACACCTTGCTCCTTTGGCGGCGTGTCTGT 
                 
                 
                   GATCACACCTGGCACCAATACAAGCAACCAGGTGGCCGTGCTGTACCAGGGCGTGAA 
                 
                 
                   TTGTACCGAGGTGCCAGTGGCCATCCACGCAGACCAGCTCACCCCCACATGGCGGGT 
                 
                 
                   GTATTCCACCGGCTCTAACGTGTTCCAGACAAGAGCCGGCTGCCTGATCGGAGCCGA 
                 
                 
                   GCATGTGAACAATTCCTACGAGTGCGACATCCCCATCGGAGCCGGCATCTGTGCCTCT 
                 
                 
                   TATCAGACCCAGACAAACAGCCCTGGCTCCGCCAGCTCCGTGGCCTCTCAGAGCATC 
                 
                 
                   ATCGCCTACACCATGTCCCTGGGCGTGGAGAACAGCGTGGCCTATTCTAACAATAGC 
                 
                 
                   ATCGCCATCCCCACCAACTTTACAATCTCTGTGACCACAGAGATCCTGCCTGTGAGCA 
                 
                 
                   TGACCAAGACATCCGTGGACTGCACAATGTACATCTGTGGCGATTCCACCGAGTGCT 
                 
                 
                   CTAACCTGCTGCTGCAGTATGGCAGCTTCTGTACCCAGCTCAACCGGGCCCTGACAGG 
                 
                 
                   AATCGCAGTGGAGCAGGACAAGAACACACAGGAGGTGTTTGCCCAGGTGAAGCAGA 
                 
                 
                   TCTACAAGACCCCACCCATCAAGGACTTCGGCGGCTTTAATTTCTCCCAGATCCTGCC 
                 
                 
                   AGATCCCTCCAAGCCATCTAAGCGGAGCTTCATCGAGGACCTGCTGTTTAACAAGGT 
                 
                 
                   GACCCTGGCCGATGCCGGCTTTATCAAGCAGTACGGCGATTGCCTGGGCGACATCGC 
                 
                 
                   AGCCAGAGACCTGATCTGTGCCCAGAAGTTCAATGGCCTGACCGTGCTGCCTCCACT 
                 
                 
                   GCTGACAGATGAGATGATCGCCCAGTATACAAGCGCCCTGCTGGCCGGCACCATCAC 
                 
                 
                   ATCCGGATGGACCTTCGGGGCCGGGGCCGCCCTGCAGATCCCCTTCGCCATGCAGAT 
                 
                 
                   GGCCTACAGGTTTAACGGCATCGGCGTGACCCAGAATGTGCTGTATGAGAACCAGAA 
                 
                 
                   GCTGATCGCCAATCAGTTCAACTCCGCCATCGGCAAGATCCAGGACAGCCTGTCTAG 
                 
                 
                   CACAGCCTCCGCCCTGGGCAAGCTGCAGGATGTGGTGAATCAGAACGCCCAGGCCCT 
                 
                 
                   GAATACCCTGGTGAAGCAGCTCAGCAGCAACTTCGGGGCCATCAGCAGCGTGCTGAA 
                 
                 
                   CGACATCCTGAGCCGGCTGGACCCCCCTGAGGCAGAGGTGCAGATCGACAGGCTGAT 
                 
                 
                   CACAGGCCGCCTGCAGTCTCTGCAGACCTACGTGACACAGCAGCTCATCAGGGCCGC 
                 
                 
                   CGAGATCAGAGCCAGCGCCAATCTGGCCGCCACCAAGATGTCCGAGTGCGTGCTGGG 
                 
                 
                   CCAGTCTAAGCGCGTGGACTTCTGTGGCAAGGGCTACCACCTGATGAGCTTTCCACA 
                 
                 
                   GTCCGCCCCCCACGGAGTGGTGTTCCTGCATGTGACCTATGTGCCTGCCCAGGAGAA 
                 
                 
                   GAACTTTACCACAGCCCCAGCCATCTGCCACGATGGCAAGGCACACTTCCCCAGGGA 
                 
                 
                   GGGCGTGTTCGTGAGCAACGGCACCCACTGGTTCGTGACACAGCGCAACTTCTACGA 
                 
                 
                   GCCTCAGATCATCACCACAGACAATACATTCGTGTCTGGCAACTGTGACGTGGTCATC 
                 
                 
                   GGCATCGTGAACAATACCGTGTACGATCCCCTGCAGCCTGAGCTGGACAGCTTCAAG 
                 
                 
                   GAGGAGCTGGATAAGTACTTCAAGAATCACACCTCCCCCGACGTGGATCTGGGCGAC 
                 
                 
                   ATCTCTGGCATCAATGCCAGCGTGGTGAACATCCAGAAGGAGATCGACAGGCTGAAC 
                 
                 
                   GAGGTGGCCAAGAATCTGAACGAGAGCCTGATCGATCTGCAGGAGCTGGGCAAGTAT 
                 
                 
                   GAGCAGTACATCAAGTTCTTTTTCATCATCGGCCTGATCATCGGCCTGTTCCTGGTGC 
                 
                 
                   TGCGCGTGGGCATCCACCTGTGCATCAAGCTGAAGCACACCAAGAAGAGGCAGATCT 
                 
                 
                   ACACAGACATCGAGATGAACCGCCTGGGCAAGTGA 
                 
                 
                     
                 
              
             
           
         
       
     
    
    
     DETAILED DESCRIPTION OF THE EMBODIMENTS 
     Coronaviruses, such as SARS-CoV, MERS-CoV and SARS-Cov-2, are enveloped viruses having an RNA genome of about 30,000 bases. They fall within the beta genus of coronaviruses. They contain a nucleocapsid surrounded by a lipid bilayer derived from the host cell. An envelope-anchored spike protein (called “S”) mediates the entry of the coronavirus into host cells by binding a host receptor and then fusing viral and host membranes. A defined RBD is the receptor for angiotensin converting enzyme 2 (ACE2). (Wan et al., J. Vir. (2020) 94: 1). Coronavirus S proteins contain three copies of an 51 subunit and three copies of an S2 subunit. Coronavirus S proteins are cleaved into 51 and S2 subunits by furin during protein biosynthesis. The two subunits trimerize and fold into a metastable prefusion conformation. The 51 subunit is responsible for receptor binding while the S2 subunit mediates membrane fusion. 
     SARS-CoV and SARS-CoV-2 spike protein share about 76% sequence homology, suggesting that these two viruses share the same receptor, ACE2. There is lower sequence similarity between SARS-CoV-2 and MERS-CoV. 
     Studies on the genomes of SARS-CoV-2 isolated from patients over the span of four months from December 2019 to March 2020 showed that the overall similarity of the human strains declined over the four month period indicating mutation of the virus had occurred within the human population to 0.988468, corresponding to an average of 348.33 nucleotide differences. Such changes imply evolutional changes of this virus, which might result in attenuation or more virulent strains (Li et al 2020. Xidan University). Subsequently, the viral variant which was predominant prior to March 2020, D614, was overtaken by another variant which has a single amino acid change to the spike protein, G614, even in areas where D614 was well established (Korber et al, (2020) Cell, 4:812-827). Subsequently, in late 2020, an unexpected rise in reported COVID-19 cases was attributed to the emergence of the new variants, Alpha (B.1.1.7) in the UK and Beta (501Y.V2) in South Africa (Fontanet et al, (2021) the Lancet, 397: 952-954). Both variants have a mutation (N501Y) in the RBD of the spike protein that is reported to contribute to increased transmission, with estimates ranging between 40% and 70% for increased transmission. The Beta variant has two additional mutations (E484K and K417N) in the spike protein that confer a potential immune escape to antibodies. A further variant of concern, Gamma, emerged in Brazil with another set of mutations (N501Y, E484K, and K417T). The Delta variant includes RBD mutation L452R which contributes to escape from neutralization by antibodies induced by previously acquired immunity and mutation P681R in the furin cleavage site which is related to an increase the rate of S1-S2 cleavage, resulting in better transmissibility. Emergence of escape mutants is a major concern because most of the licensed vaccines against COVID-19 are based on the sequence of spike protein from the ancestral Wuhan strain of SARS-CoV-2. 
     An important concern is whether the currently available COVID-19 vaccines will be able to protect against infection or disease from the SARS-CoV-2 variants. Preliminary research suggests sera from individuals immunized with the mRNA COVID-19 vaccines neutralized a pseudovirus analogous to the U.K. variant but were less effective against a pseudovirus analogous to the South Africa variant (Yang et al (2021) Nature, doi.org/10.1038/s41586-021-03324-6). Moreover, preliminary results of studies using viral vector vaccines demonstrated good efficacy against the UK variant but poor efficacy against the South Africa variant (Madhi et al (2021) N.E.J.M. DOI: 10.1056/NEJMoa2102214). Therefore, it appears that a vaccine which is capable of inducing production of broadly reactive antibodies would be required to provide protection from infection by variant strains of coronavirus which include multiple mutations. 
     The inventors herein have made vaccines against beta coronavirus which comprises a VLP. VLPs are multiprotein structures which are generally composed of one or more viral proteins. VLP&#39;s mimic the conformation of viruses but lack genetic material, and therefore are not infectious. They can form (or “self-assemble”) upon expression of a viral structural protein under appropriate circumstances. VLP vaccines overcome some of the disadvantages of more traditional vaccines prepared using attenuated viruses because they can be produced without the need to have any live virus present during the production process. A wide variety of VLPs have been prepared. For example, VLPs including single or multiple capsid proteins either with or without envelope proteins and/or surface glycoproteins have been prepared. In some cases, VLPs are non-enveloped and assemble by expression of just one major capsid protein. In other cases, VLPs are enveloped and can comprise multiple antigenic proteins found in the corresponding native virus. Self-assembly of enveloped VLPs is more complex than non-enveloped VLPs because of the complex reactions required for fusion with the host cell membrane (Garrone et al., 2011 Science Trans. Med. 3: 1-8) and “budding” of the VLP to form a fully enveloped separate particle. Accordingly, self-assembly of enveloped VLPs may not be successful and the formation and stability of enveloped VLP particles is difficult to predict. Formation of intact VLPs can be confirmed by imaging of the particles using electron microscopy. 
     VLPs typically resemble their corresponding native virus and can be multivalent particulate structures. The present disclosure encompasses the recognition that presentation of surface glycoproteins in the context of a VLP is advantageous for induction of neutralizing antibodies against such polypeptide as compared to other forms of antigen presentation, e.g., soluble antigens not associated with a VLP. Neutralizing antibodies most often recognize tertiary or quaternary structures; this often requires presenting antigenic proteins, like envelope glycoproteins, in their native viral conformation. VLP&#39;s present epitopes in a highly structured, repetitive array that enables efficient crosslinking of B cell receptors, leading to activation and expansion of high affinity B cells and subsequent antibody production (Bachmann, 1993). Indeed, VLP expression of a B cell antigen improved neutralizing titers by over 10-fold relative to immunization with the same amount of recombinant protein (Kirchmeier, 2014). Accordingly, use of VLPs as a vaccine modality may expand higher affinity B cell repertoires relative to recombinant protein or DNA/mRNA-based approaches, the latter approach being used in two widely used COVID-19 vaccines. 
     The VLPs of the invention comprise retroviral vectors. Retroviruses are enveloped RNA viruses that belong to the family Retroviridae. After infection of a host cell by a retrovirus, RNA is transcribed into DNA via the enzyme reverse transcriptase. DNA is then incorporated into the host cell&#39;s genome by an integrase enzyme and thereafter replicates as part of the host cell&#39;s DNA. The Retroviridae family includes the following genera Alpharetrovirus, Betaretrovirus, Gammearetrovirus, Deltaretrovirus, Epsilonretrovirus, Lentivirus and Spumavirus. The hosts for this family of retroviruses generally are vertebrates. Retroviruses produce an infectious virion containing a spherical nucleocapsid (the viral genome in complex with viral structural proteins) surrounded by a lipid bilayer derived from the host cell membrane. 
     Retroviral vectors can be used to generate VLPs that lack a retrovirus-derived genome and are therefore non-replicating. This is accomplished by replacement of most of the coding regions of the retrovirus with genes or nucleotide sequences to be transferred; so that the vector is incapable of making proteins required for additional rounds of replication. Depending on the properties of the glycoproteins present on the surface of the particles, VLPs have limited ability to bind to and enter the host cell but cannot propagate. Therefore, VLPs can be administered safely as an immunogenic composition (e.g., a vaccine). 
     The present invention utilizes VLPs comprising one or more retroviral structural proteins. In some embodiments, a structural protein for use in accordance with the present invention is Alpharetrovirus (e.g., Avian Leukosis Virus), Betaretrovirus (Mouse Mammary Tumor Virus), Gammearetrovirus (Murine Leukemia Virus), Deltaretrovirus (Bovine Leukemia Virus), Epsilonretrovirus (Walley Dermal Sarcoma Virus), Lentivirus (Human Immunodeficiency Virus 1) or Spumavirus (Chimpanzee Foamy Virus) structural protein. In certain embodiments, a structural polyprotein is a Murine Leukemia Virus (MLV) structural protein. In an embodiment of the invention the structural protein in a Moloney Murine Leukemia Virus (MMLV). Genomes of these retroviruses are readily available in databases. 
     In some embodiments, the retroviral structural protein for use in accordance with the present invention is a Gag polypeptide. The Gag proteins of retroviruses have an overall structural similarity and, within each group of retroviruses, are conserved at the amino acid level. Retroviral Gag proteins primarily function in viral assembly. Expression of Gag of some viruses (e.g., murine leukemia viruses, such as MMLV) in some host cells, can result in self-assembly of the expression product into VLPs. The Gag gene expression product in the form of a polyprotein gives rise to the core structural proteins of the VLP. Functionally, the Gag polyprotein is divided into three domains: the membrane binding domain, which targets the Gag polyprotein to the cellular membrane; the interaction domain which promotes Gag polymerization; and the late domain which facilitates release of nascent virions from the host cell. In general, the form of the Gag protein that mediates viral particle assembly is the polyprotein. Retroviruses assemble an immature capsid composed of the Gag polyprotein but devoid of other viral elements like viral protease with Gag as the structural protein of the immature virus particle. 
     A suitable Gag polypeptide for use in the invention is substantially homologous to a known retroviral Gag polypeptide. The MMLV-Gag gene encodes a 65 kDa polyprotein precursor which is proteolytically cleaved into 4 structural proteins (Matrix (MA); p12; Capsid (CA); and Nucleocapsid (NC)), by MLV protease, in the mature virion. In the absence of MLV protease, the polyprotein remains uncleaved and the resulting particle remains in an immature form. The morphology of the immature particle is different from that of the mature particle. In some embodiments of the invention, the Gag sequence does not include a gene encoding MLV protease. The gene encoding the MMLV nucleic acid is SEQ ID NO: 2. An exemplary codon optimized sequence of MMLV nucleic acid is provided as SEQ ID NO: 3. 
     Therefore, in some embodiments, a Gag polypeptide suitable for the present invention is substantially homologous to an MMLV-Gag polypeptide (SEQ ID NO:1). In some embodiments, a Gag polypeptide suitable for the present invention has an amino acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO:1. In some embodiments, a Gag polypeptide suitable for the present invention is substantially identical to, or identical to SEQ ID NO: 1. 
     In some embodiments, a suitable MMLV-Gag polypeptide is encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:2. In some embodiments, a suitable MMLV-Gag polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 2 or a codon degenerate version thereof. 
     As is well known to those of skill in the art, it is possible to improve the expression of a nucleic acid sequence in a host organism by replacing the nucleic acids coding for a particular amino acid (i.e. a codon) with another codon which is better expressed in the host organism. One reason that this effect arises is due to the fact that different organisms show preferences for different codons. The process of altering a nucleic acid sequence to achieve better expression based on codon preference is called codon optimization. Various methods are known in the art to analyze codon use bias in various organisms and many computer algorithms have been developed to implement these analyses in the design of codon optimized gene sequences. Therefore, in some embodiments, a suitable MMLV-Gag polypeptide is encoded by a codon optimized version of a nucleic acid sequence encoding MMLV-Gag and having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:3. In some embodiments, a suitable MMLV-Gag polypeptide is encoded by a nucleic acid sequence which is substantially identical to, or identical to, SEQ ID NO: 3. 
     As is well known in this art, amino acid or nucleic acid sequences may be compared using any of a variety of algorithms, including those available in commercial computer programs such as BLASTN for nucleotide sequences and BLASTP, gapped BLAST, and PSI-BLAST for amino acid sequences. Examples of such programs are described in Altschul, et al., 1990 , J. Mol. Biol.,  215(3): 403-410; Altschul, et al., 1996 , Methods in Enzymology  266:460-480; Altschul, et al., 1997 Nucleic Acids Res. 25:3389-3402; Baxevanis, et al., 1998 , Bioinformatics: A Practical Guide to the Analysis of Genes and Proteins , Wiley; and Misener, et al., (eds.), 1999 , Bioinformatics Methods and Protocols  (Methods in Molecular Biology, Vol. 132), Humana Press. In addition to identifying homologous sequences, the programs mentioned above typically provide an indication of the degree of homology. In some embodiments, two sequences are considered to be substantially homologous if at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more of their corresponding residues are homologous over a relevant stretch of residues. In some embodiments, the relevant stretch is a complete sequence. In some embodiments, the relevant stretch is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or more residues. 
     Alternatively, the Gag polypeptide used in the invention may be a modified retroviral Gag polypeptide containing one or more amino acid substitutions, deletions, and/or insertions as compared to a wild-type or naturally-occurring Gag polypeptide while retaining substantial self-assembly activity. Typically, in nature, a Gag protein includes a large C-terminal extension which may contain retroviral protease, reverse transcriptase, and integrase enzymatic activity. Assembly of VLPs, however, generally does not require the presence of such components. In some cases, a retroviral Gag protein alone (e.g., lacking a C-terminal extension, lacking one or more of genomic RNA, reverse transcriptase, viral protease, or envelope protein) can self-assemble to form VLPs both in vitro and in vivo (Sharma S et al., 1997, Proc. Natl. Acad. Sci. USA 94: 10803-8). 
     The inventors of the present application have made VLPs which express beta coronavirus envelope glycoproteins on the surface which can cause an immune response in a subject. A humoral immune response is an immune response mediated by antibody molecules. Certain antibodies, called neutralizing antibodies, defend cells from infection by a virus and associated biological effects by recognizing and binding to a particular protein or antigen expressed by the virus. The envelope protein of coronaviruses are important targets for production of neutralizing antibodies. It is well known to those in the art that retroviral Gag-based enveloped VLPs can be used to express a variety of envelope glycoproteins for the purpose of eliciting neutralizing antibody responses. More specifically, evidence exists for expression of Class I viral fusion proteins such as HIV-1 gp120, metapneumovirus and Influenza HA, as well as Class III fusion proteins such as VSV G protein and CMV gB protein (Mammano et al., 1997, J. Virol. 71:3341-3345; Levy et al., 2013, Vaccine 31:2778-2785; Lemaitre et al., 2011, Clin. Microbiol. Infect. 1:732-737; Garrone et al, 2011; Kirchmeier et al., 2014, CVI 21: 174-180). However, there is little known about expression of coronavirus spike proteins, particularly with MLV-derived Gag. In U.S. Pat. No. 8,920,812, Example 1 describes a failure to express RSV F glycoprotein, a class II viral fusion protein, on the surface of a VLP produced using MLV Gag. The inventor hypothesized that the presence of the RSV F glycoprotein interfered with budding of the Gag viral particle through the cell membrane (see column 41, line 50). It was therefore not predictable that a retroviral Gag-based enveloped virus-like particle could be used to successfully express the coronavirus spike protein. Nevertheless, the present inventors have made several different embodiments of a beta coronavirus vaccine comprising one or more spike polypeptide antigens (e.g., from SARS CoV-2, SARS CoV and MERS-CoV) on the surface of a VLP. In some embodiments, the spike polypeptide antigens comprise modified polypeptides. In some embodiments, the spike polypeptide antigens have more than one genetic modification. 
     In some embodiments, a VLP of the invention includes a fusion protein of a spike polypeptide from a beta coronavirus (e.g., all or part of an extracellular portion of an beta coronavirus spike polypeptide) and a transmembrane and/or cytoplasmic domain that is not found in nature in the beta coronavirus protein (e.g., from another virus). In some embodiments, a fusion protein includes a spike polypeptide from a beta coronavirus (e.g., all or part of an extracellular portion of the spike polypeptide) and a transmembrane domain and/or cytoplasmic domain found in nature in the glycoprotein G from VSV which is referred to as VSV-G. The nucleotide and amino acid sequences of the VSV-G protein are known in the art. 
     The transmembrane domain of VSV-G can function to target the viral glycoprotein to the cell membrane (Compton T et al., 1989, Proc Natl Acad Sci USA 86:4112-4116). Swapping the transmembrane and cytoplasmic domains of VSV-G for the transmembrane and cytoplasmic domains of another protein has been used to increase the yield of the protein of interest in the VLP preparation and increase immunogenicity to neutralizing antibody response (Garrone et al., 2011). This modification was successful to increase yield and activity of a VLP expressing HCV-E1 protein (Garrone et al, 2011) and CMV-gB protein (Kirchmeier et al, 2014). However, this modification has also been associated with a significant loss of immunogenicity when used with certain viral antigens. In addition, expression of some glycoproteins has decreased after replacement of the transmembrane/cytoplasmic domain of the antigenic glycoprotein with the transmembrane/cytoplasmic domain from VSV. For example, loss of glycoprotein was reported in SARS virus (Broer et al., 2006, J. Vir. 80, 1302-1310). In RSV, a significant loss of immunogenicity was observed when the antigenic surface protein was modified to replace the transmembrane component with a sequence from VSV. 
     In some embodiments, an immunogenic composition of the present invention comprises a VLP comprising a wild type spike polypeptide from SARS-CoV-2, the sequence of which is SEQ ID NO: 4 or a codon degenerate version of SEQ ID NO: 4. A nucleic acid which encodes for the polypeptide is shown as SEQ ID NO: 5. A codon optimized version of SEQ ID NO: 5 is shown as SEQ ID NO: 6. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 4. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 4 or a codon degenerate version of SEQ ID NO: 4. In some embodiments, the polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 5. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 5, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 6. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 6. 
     In some embodiments, an immunogenic composition of the present invention comprises a VLP comprising a modified spike polypeptide from SARS-CoV-2 which has been modified to replace the transmembrane and cytoplasmic segments with corresponding segments from VSV, the sequence of which is SEQ ID NO: 26 or a codon degenerate version of SEQ ID NO: 26. A nucleic acid which encodes for the polypeptide is shown as SEQ ID NO: 25. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 25. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 25 or a codon degenerate version of SEQ ID NO: 25. In some embodiments, the polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 26. In some embodiments, the mutated polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 26. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 26, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 27. 
     In some embodiments, an immunogenic composition of the present invention comprises a VLP comprising a wild type spike polypeptide from SARS-CoV, the sequence of which is SEQ ID NO: 7 or a codon degenerate version of SEQ ID NO: 7. A nucleic acid which encodes for the polypeptide is shown as SEQ ID NO: 8. A codon optimized version of SEQ ID NO: 8 is shown as SEQ ID NO: 9. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 7. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 7 or a codon degenerate version of SEQ ID NO: 7. In some embodiments, the polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 8. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 8, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 9. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 9. 
     In some embodiments, an immunogenic composition of the present invention comprises a VLP comprising a wild type spike polypeptide from MERS-CoV, the sequence of which is SEQ ID NO: 10 or a codon degenerate version of SEQ ID NO: 10. A nucleic acid which encodes for the polypeptide is shown as SEQ ID NO: 11. A codon optimized version of SEQ ID NO:11 is shown as SEQ ID NO: 12. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 10. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 10 or a codon degenerate version of SEQ ID NO: 10. In some embodiments, the polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 11. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 11, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 12. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 12. 
     In some embodiments, immunogenic compositions of the present invention comprise VLPs comprising variants of beta coronavirus spike glycoproteins. In some embodiments, a variant spike glycoprotein has been modified to delete the furin cleavage site from the spike polypeptide. In some embodiments, the spike glycoprotein has been modified to replace lysine (986) and valine (987) residues with proline residues. In some embodiments, the spike glycoprotein has been modified to delete the furin cleavage site and to replace lysine (986) and valine (987) residues with proline residues. Each such modification is further described below. 
     It is known that the coronavirus spike protein includes a site where the protease, furin, cleaves the S polypeptide into 51 and S2 subunits during the process of virion maturation. A modified spike protein construct was produced wherein the amino acid sequence was modified to remove the furin cleavage site, thus retaining the spike polypeptide in its immature form. It is possible that the furin-cleavage site mutated version of the spike protein, which does not undergo normal cleavage and maturation, will show enhanced cell receptor binding and cell entry, indicating that immunity against this structure may result in humoral immunity with greater neutralizing activity. 
     In some embodiments, an immunogenic composition of the invention comprises a VLP comprising a modified SARS-CoV-2 spike polypeptide with a mutated furin cleavage site as compared to a wild-type or naturally-occurring SARS-CoV-2 spike polypeptide. The sequence for an exemplary modified SARS-CoV-2 polypeptide is shown as SEQ ID NO: 16. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 16. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 16 or a codon degenerate version of SEQ ID NO: 16. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 17. In some embodiments, the modified polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 17, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 18. In some embodiments, the mutated polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 18. 
     It is known from previous studies of SARS-CoV and MERS-CoV that substitution of two amino acid residues with proline residues results in stabilisation of the S2 subunit in its prefusion conformation (Pallesen et al., 2017 PNAS. 114:35; Wrapp et al (2020) Science: 367: 1260-1263). Therefore, it is possible that such a mutation could enhance the immune response to coronavirus. Accordingly, a SARS-CoV-2 polypeptide construct was prepared which has been modified to replace lysine (986) and valine (987) residues with proline residues. In some embodiments, an immunogenic composition of the invention comprises a VLP comprising a SARS-CoV-2 polypeptide which has been modified to replace lysine (986) and valine (987) residues with proline residues as compared to a wild-type or naturally-occurring SARS-CoV-2 polypeptide. The sequence of an exemplary modified polypeptide is shown in SEQ ID NO: 13. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 13. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 13 or a codon degenerate version of SEQ ID NO: 13. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 14. In some embodiments, the modified polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 14, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO:15. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 15. 
     In a further variation, a SARS-CoV-2 polypeptide construct was prepared which has been modified to replace lysine (986) and valine (987) residues with proline residues and which have been further modified to remove the furin cleavage site. In some embodiments, an immunogenic composition of the invention comprises a VLP comprising a SARS-CoV-2 polypeptide which has been modified to replace lysine (986) and valine (987) residues with proline residues and to remove the furin cleavage site as compared to a wild-type or naturally-occurring SARS-CoV-2 polypeptide. The sequence of an exemplary modified polypeptide is shown in SEQ ID NO: 19. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 19. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is SEQ ID NO: 19 or a codon degenerate version of SEQ ID NO: 19. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 20. In some embodiments, the modified polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 20, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO:21. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 21. 
     In some embodiments, a VLP described herein comprises a fusion protein comprising an extracellular domain (or a portion thereof) of a coronavirus spike polypeptide, and a transmembrane domain and cytoplasmic tail from an envelope protein from VSV. In some embodiments, the immunogenic composition of the invention comprises a VLP comprising a coronavirus spike polypeptide modified to replace the transmembrane domain and cytoplasmic tail with the transmembrane domain and cytoplasmic tail from VSV. Any of the coronavirus spike proteins described herein may be modified to replace the transmembrane domain and cytoplasmic tail with a transmembrane domain and cytoplasmic tail from VSV. 
     In one particular embodiment, the inventors have constructed a SARS-CoV-2 spike protein which protein has been modified to replace the transmembrane domain and cytoplasmic tail with a transmembrane domain and cytoplasmic tail from VSV; to replace lysine (986) and valine (987) residues with proline residues; and to remove the furin cleavage site. This triple modified SARS-CoV-2 protein includes the double proline mutation directed to enhanced stability and a mutated furin cleavage site, which is associated with enhanced receptor binding. Further, it includes the transmembrane domain and cytoplasmic tail from VSV, which are associated with improved expression on the VLP envelope. The sequence of this triple modified coronavirus spike polypeptide is shown as SEQ ID NO: 22 (shown above with the portion from VSV in bold text at the end of the sequence). In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 22. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 23. In some embodiments, the modified polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 23, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 23. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 24. 
     In a further embodiment, the inventors have constructed a SARS-CoV-2 spike protein from the Beta variant which protein has been modified to replace the transmembrane domain and cytoplasmic tail with a transmembrane domain and cytoplasmic tail from VSV; to replace lysine (986) and valine (987) residues (based on the positions in the Wuhan reference) with proline residues; and to remove the furin cleavage site. This triple modified SARS-CoV-2 protein from the Beta variant includes the double proline mutation directed to enhanced stability and a mutated furin cleavage site, which is associated with enhanced receptor binding. Further, it includes the transmembrane domain and cytoplasmic tail from VSV, which are associated with improved expression on the VLP envelope. The sequence of this triple modified coronavirus spike polypeptide is shown as SEQ ID NO: 28. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising a polypeptide having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 28. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 29. In some embodiments, the modified polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 29, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 29. In some embodiments, the modified polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 30. 
     Mixtures of antigens can induce broad reactive immunity therefore, combinations of coronavirus antigens can be used to enhance the breadth of the immune response. VLPs can be used to express two (bivalent) or three (trivalent) viral antigens in their native conformation, thus inducing a potent B cell response. Previous studies using Zika viral epitopes demonstrated that the combination of two antigens on a single bivalent VLP generated a significantly more potent immune response than two monovalent VLPs expressing the same antigens (U.S. Pat. No. 8,920,812). 
     Accordingly, the VLPs of the present disclosure include bivalent VLPs containing two wild type coronavirus spike proteins, two modified coronavirus spike proteins described herein or any combination of the wild type and modified coronavirus spike proteins described herein. The VLPs of the present disclosure also include trivalent VLPs containing three wild type coronavirus spike proteins, three modified coronavirus spike proteins described herein or any combination of the wild type and modified coronavirus spike proteins described herein. One or more of any of the wild type or modified spike proteins expressed on a bivalent or a trivalent VLP may be further modified to replace the transmembrane domain and the cytoplasmic tail with the transmembrane domain and cytoplasmic tail from VSV. 
     In a preferred embodiment, the VLP of the present disclosure is a trivalent VLP comprising a spike protein from SARS-CoV-2, a spike protein from SARS-CoV and a spike protein from MERS-CoV. One or more of the spike proteins may be modified to replace the transmembrane domain and the cytoplasmic tail with the transmembrane domain and cytoplasmic tail from VSV. 
     In some embodiments, an immunogenic composition of the present invention comprises a trivalent VLP comprising a wild type spike polypeptide from SARS-CoV-2, the sequence of which is SEQ ID NO: 4 or a codon degenerate version of SEQ ID NO: 4; a spike polypeptide from SARS-CoV the sequence of which is SEQ ID NO: 7 or a codon degenerate version of SEQ ID NO: 7; and a spike polypeptide from MERS the sequence of which is SEQ ID NO: 10 or a codon degenerate version of SEQ ID NO: 10. A nucleic acid which encodes for the SARS-CoV-2 polypeptide is shown as SEQ ID NO: 5. A codon optimized version of SEQ ID NO: 5 is shown as SEQ ID NO: 6. A nucleic acid which encodes for the SARS-CoV polypeptide is shown as SEQ ID NO: 8. A codon optimized version of SEQ ID NO: 8 is shown as SEQ ID NO: 9. A nucleic acid which encodes for the MERS polypeptide is shown as SEQ ID NO: 11. A codon optimized version of SEQ ID NO:11 is shown as SEQ ID NO: 12. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising polypeptides having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 4, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 7 and 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 10. In some embodiments, the SARS-CoV-2 polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 5. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 5, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 6. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 6. In some embodiments, the SAR-CoV polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 8. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 8, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 9. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 9. In some embodiments, the MERS polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 11. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 11, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 12. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 12. 
     In some embodiments, an immunogenic composition of the present invention comprises a trivalent VLP comprising a modified spike polypeptide from SARS-CoV-2, the sequence of which is SEQ ID NO: 22 or a codon degenerate version of SEQ ID NO: 22; a spike polypeptide from SARS-CoV the sequence of which is SEQ ID NO: 7 or a codon degenerate version of SEQ ID NO: 7; and a spike polypeptide from MERS the sequence of which is SEQ ID NO: 10 or a codon degenerate version of SEQ ID NO: 10. A nucleic acid which encodes for the SARS-CoV-2 polypeptide is shown as SEQ ID NO: 5. A codon optimized version of SEQ ID NO: 5 is shown as SEQ ID NO: 6. A nucleic acid which encodes for the SARS-CoV polypeptide is shown as SEQ ID NO: 8. A codon optimized version of SEQ ID NO: 8 is shown as SEQ ID NO: 9. A nucleic acid which encodes for the MERS polypeptide is shown as SEQ ID NO: 11. A codon optimized version of SEQ ID NO:11 is shown as SEQ ID NO: 12. In some embodiments, the present invention comprises an immunogenic composition comprising a VLP comprising polypeptides having an amino acid sequence which is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 22, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 7 and 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 10. In some embodiments, the SARS-CoV-2 polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 23. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 23, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 24. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 24. In some embodiments, the SAR-CoV polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 8. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 8, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 9. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 9. In some embodiments, the MERS polypeptide is encoded by a nucleic acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 11. In some embodiments, the polypeptide is encoded by a codon optimized version of the nucleic acid sequence of SEQ ID NO: 11, which is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to the SEQ ID NO: 12. In some embodiments, the polypeptide is encoded by a nucleic acid sequence having SEQ ID NO: 12. 
     As can be seen in the Examples, the VLPs of the invention were able to elicit a strong immune response to SARS-CoV-2. In particular, each of the modified spike variants described herein was effective to induce a strong immune response (see Example 5). The trivalent VLPs of the invention (see Example 6) induced an antibody response against SARS-Cov-2, SARS-CoV and MERS. Moreover, immunization with the trivalent VLPs of the invention induced antibodies that recognized a related seasonal human coronavirus, 0C43, not included within the vaccine, demonstrating that the trivalent VLP has an ability to broaden immunity against coronaviruses. Unexpectedly, relative to immunization with a monovalent VLP, trivalent VLPs enriched the induction of antibodies with functional, neutralizing activity. This enrichment of neutralizing antibodies in shown in Table 8, which shows the ratio of endpoint neutralizing antibody titer to the endpoint antibody binding titer, using sera obtained from vaccinated mice. 
     The monovalent VLP which expresses the triple modified SARS-CoV-2 spike protein provides significant protection against infection by SARS-CoV-2 as demonstrated by a challenge study in golden hamster (Example 7). As shown in Example 7, the hamsters which were vaccinated with the VLP had significantly lower levels of viral RNA and improved clinical presentation as shown by body weight. Furthermore, the immunized hamsters were able to mount a stronger cytokine response than the unvaccinated hamsters. Interim data from a Phase 1a clinical trial in humans using a vaccine formulation comprising the triple modified SARS-CoV-2 spike protein adjuvanted with aluminum phosphate adjuvant showed that all subjects receiving two doses had seroconverted 56 days after the second dose (see Example 11). 
     The VLPs of the invention have demonstrated a broad immune response that is effective against a variant of SARS-CoV-2. As described in Example 9, a trivalent VLP expressing the triple modified SARS-CoV-2 spike protein, a native MERS spike protein and a native SARS-CoV protein and a monovalent VLP expressing the triple modified SARS-CoV-2 spike protein were evaluated for their ability to induce antibodies against the Beta (South Africa) variant of SARS-CoV-2 in mice. Surprisingly, both the trivalent and the monovalent constructs elicited antibodies to the Beta variant. Even more surprising was the fact that the antibody titres were similar for the Beta strain and the ancestral L strain of SARS-CoV-2. Accordingly, both the trivalent and the monovalent VLPs expressing the triple modified SARS-CoV-2 spike protein were unexpectedly effective at inducing a potent antibody response to a SARS-CoV-2 variant which has demonstrated significant escape from other COVID-19 vaccines. 
     Enhanced antibody binding and neutralizing activity was observed when a monovalent VLP which expresses a triple modified SARS-CoV-2 spike protein based on the Beta variant (SEQ ID NO: 30) was used as a booster vaccine following a first vaccination with a vaccine based on a triple modified SARS-CoV-2 spike protein based on the ancestral strain (SEQ ID NO: 24). This heterologous boosting strategy also resulted in enhanced antibody binding against the Delta variant indicating that the combination strategy enhances the breadth of the humoral immune response (see Example 12). 
     The VLPs of the invention also had an effect on the nature of the antibody response. As shown in Example 10, mice immunized with a monovalent VLP of the invention expressing wild type SARS-CoV-2 spike protein produced a higher amount of IgG2b than those immunized with a recombinant spike protein. Higher IgG2b is associated with a TH1 immune response and may result in a higher level of cell-mediated immunity. Therefore, presentation of the spike protein on an VLP resulted in a response correlated to cell-mediated immunity. 
     It will be appreciated that a composition comprising VLPs will typically include a mixture of VLPs with a range of sizes. It is to be understood that the diameter values listed below correspond to the most frequent diameter within the mixture. In some embodiments &gt;90% of the vesicles in a composition will have a diameter which lies within 50% of the most frequent value (e.g., 1000±500 nm). In some embodiments, the distribution may be narrower, e.g., &gt;90% of the vesicles in a composition may have a diameter which lies within 40, 30, 20, 10 or 5% of the most frequent value. In some embodiments, sonication or ultra-sonication may be used to facilitate VLP formation and/or to alter VLP size. In some embodiments, filtration, dialysis and/or centrifugation may be used to adjust the VLP size distribution. 
     In general, VLPs produced in accordance with the methods of the present disclosure may be of any size. In certain embodiments, the composition may include VLPs with diameters in the range of about 20 nm to about 300 nm. In some embodiments, a VLP is characterized in that it has a diameter within a range bounded by a lower limit of 20, 30, 40, 50, 60, 70, 80, 90, or 100 nm and bounded by an upper limit of 300, 290, 280, 270, 260, 250, 240, 230, 220, 210, 200, 190, 180, or 170 nm. In some embodiments, VLPs within a population show an average diameter within a range bounded by a lower limit of 20, 30, 40, 50, 60, 70, 80, 90, or 100 nm and bounded by an upper limit of 300, 290, 280, 270, 260, 250, 240, 230, 220, 210, 200, 190, 180, or 170 nm. In some embodiments, VLPs in a population have a polydispersity index that is less than 0.5 (e.g., less than 0.45, less than 0.4, or less than 0.3). In some embodiments, VLP diameter is determined by nanosizing. In some embodiments, VLP diameter is determined by electron microscopy. 
     VLPs in accordance with the present invention may be prepared according to general methodologies known to the skilled person. For example, nucleic acid molecules, reconstituted vectors or plasmids may be prepared using sequences which are known in the art. Such sequences are available from banks, and material may be obtained from various collections, published plasmids, etc. These elements can be isolated and manipulated using techniques well known to the skilled artisan, or available in the art. Various synthetic or artificial sequences may also be produced from computer analysis or through (high throughput) screening of libraries. Recombinant expression of the polypeptides for VLPs requires construction of an expression vector containing a polynucleotide that encodes one or more polypeptide(s). Once a polynucleotide encoding one or more polypeptides has been obtained, the vector for production of the polypeptide may be produced by recombinant DNA technology using techniques known in the art. Expression vectors that may be utilized in accordance with the present invention include, but are not limited to mammalian and avian expression vectors, bacculovirus expression vectors, plant expression vectors (e.g., Cauliflower Mosaic Virus (CaMV), Tobacco Mosaic Virus (TMV)), plasmid expression vectors (e.g., Ti plasmid), among others. 
     The VLPs of the invention may be produced in any available protein expression system. Typically, the expression vector is transferred to a host cell by conventional techniques and the transfected cells are then cultured by conventional techniques to produce VLPs. In some embodiments, VLPs are produced using transient transfection of cells. In some embodiments, VLPs are produced using stably transfected cells. Typical cell lines that may be utilized for VLP production include, but are not limited to, mammalian cell lines such as human embryonic kidney (HEK) 293, WI 38, Chinese hamster ovary (CHO), monkey kidney (COS), HT1080, C10, HeLa, baby hamster kidney (BHK), 3T3, C127, CV-1, HaK, NS/O, and L-929 cells. Specific non-limiting examples include, but are not limited to, BALB/c mouse myeloma line (NSO/1, ECACC No: 85110503); human retinoblasts (PER.C6 (CruCell, Leiden, The Netherlands)); monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol., 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells +/−DHFR (CHO, Urlaub and Chasin, Proc. Natl. Acad. Sci. USA, 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod., 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1 587); human cervical carcinoma cells (HeLa, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci., 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2). In some embodiments, cell lines that may be utilized for VLP production include insect (e.g., Sf-9, Sf-21, Tn-368, Hi5) or plant (e.g., Leguminosa, cereal, or tobacco) cells. It will be appreciated in some embodiments, particularly when glycosylation is important for protein function, mammalian cells are preferable for protein expression and/or VLP production (see, e.g., Roldao A et al., 2010 Expt Rev Vaccines 9:1149-76). 
     It will be appreciated that a cell strain may be chosen which modulates the expression of the inserted sequences, or modifies and processes the gene product in a specific way. Such modifications (e.g., glycosylation) and processing (e.g., cleavage or transport to the membrane) of protein products may be important for generation of a VLP or function of a VLP polypeptide or additional polypeptide (e.g., an adjuvant or additional antigen). Different cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed. Generally, eukaryotic host cells (also referred to as packaging cells (e.g., 293T human embryo kidney cells)) which possess appropriate cellular machinery for proper processing of the primary transcript, glycosylation and phosphorylation of the gene product may be used in accordance with the present invention. 
     VLPs may be purified according to known techniques, such as centrifugation, gradients, sucrose-gradient ultracentrifugation, tangential flow filtration and chromatography (e.g., ion exchange (anion and cation), affinity and sizing column chromatography), or differential solubility, among others. Alternatively, or additionally, cell supernatant may be used directly, with no purification step. Additional entities, such as additional antigens or adjuvants may be added to purified VLPs. 
     In accordance with the present invention, cells may be transfected with a single expression vector. In some embodiments, a single expression vector encodes more than one element of a VLP (e.g., more than one of structural polyprotein, coronavirus spike protein, etc.). For example, in some embodiments, a single expression vector encodes two or more elements of a VLP. In some embodiments, a single expression vector encodes three of more elements of a VLP. In an embodiment of the invention, a single expression vector encodes a Gag polypeptide and a coronavirus spike glycoprotein. 
     In some embodiments, cells are transfected with two or more expression vectors. For example, in some embodiments, cells are transfected with a first vector encoding a Gag polypeptide and a second vector encoding a coronavirus spike glycoprotein and “monovalent” VLPs comprising a coronavirus spike glycoprotein are produced. In some embodiments, cells are transfected with a first vector encoding a Gag polypeptide, a second vector encoding a coronavirus spike glycoprotein and a third vector encoding another coronavirus spike glycoprotein. In such embodiments, “bivalent” VLPs comprising two coronavirus spike glycoproteins are produced. In some embodiments, cells are transfected with a first vector encoding a Gag polypeptide, a second vector encoding a coronavirus spike glycoprotein, and a third vector encoding two coronavirus spike glycoproteins. In such embodiments, “trivalent” VLPs comprising three coronavirus spike glycoproteins are produced. 
     As further demonstrated in the Examples, modification of the SARS-CoV-2 spike protein had a significant effect on the yield of the VLPs. Referring to Table 1, in Example 3, the VLP expressing the triple modified SARS-CoV-2 spike protein (Group 3) showed significantly higher spike protein yield that other monovalent VLPs expressing SARS-CoV-2 spike proteins. Accordingly, this embodiment of the VLP can be manufactured in higher volumes, which is important for addressing demand in pandemic situations. 
     In some embodiments, monovalent, bivalent, or trivalent VLPs are admixed. For example, in some embodiments, monovalent and bivalent VLPs are admixed to form a trivalent VLP mixture. In some embodiments two monovalent VLPs are admixed to form a bivalent VLP mixture. 
     The present invention provides pharmaceutical compositions comprising the VLPs described herein and, optionally, further comprising the glycoproteins and glycoprotein variants described herein. In some embodiments, the present invention provides a VLP and at least one pharmaceutically acceptable excipient, adjuvant and/or carrier. Such pharmaceutical compositions may optionally comprise and/or be administered in combination with one or more additional therapeutically active substances. The provided pharmaceutical compositions are useful as prophylactic agents (i.e., vaccines) in the prevention of SARS, MERS or COVID-19 infection or of negative ramifications associated or correlated with SARS, MERS or COVID-19 infection. The provided pharmaceutical compositions are also useful as prophylactic agents against certain variants of SARS-CoV-2. In some embodiments, pharmaceutical compositions are formulated for administration to humans. 
     Pharmaceutical compositions provided here may be provided in a sterile injectable form (e.g., a form that is suitable for subcutaneous injection or intravenous infusion). For example, in some embodiments, pharmaceutical compositions are provided in a liquid dosage form that is suitable for injection. In some embodiments, pharmaceutical compositions are provided as powders (e.g. lyophilized and/or sterilized), optionally under vacuum, which are reconstituted with an aqueous diluent (e.g., water, buffer, salt solution, etc.) prior to injection. In some embodiments, pharmaceutical compositions are diluted and/or reconstituted in water, sodium chloride solution, sodium acetate solution, benzyl alcohol solution, phosphate buffered saline, etc. In some embodiments, powder should be mixed gently with the aqueous diluent (e.g., not shaken). 
     In some embodiments, provided pharmaceutical compositions comprise one or more pharmaceutically acceptable excipients (e.g., preservative, inert diluent, dispersing agent, surface active agent and/or emulsifier, buffering agent, etc.). Suitable excipients include, for example, water, saline, dextrose, sucrose, trehalose, glycerol, ethanol, or similar, and combinations thereof. Remington&#39;s The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro, (Lippincott, Williams &amp; Wilkins, Baltimore, Md., 2006) discloses various excipients used in formulating pharmaceutical compositions and known techniques for the preparation thereof. Except insofar as any conventional excipient medium is incompatible with a substance or its derivatives, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition, its use is contemplated to be within the scope of this invention. In some embodiments, pharmaceutical compositions comprise one or more preservatives. In some embodiments, pharmaceutical compositions comprise no preservative. 
     In some embodiments, a pharmaceutical composition is sufficiently immunogenic as a vaccine (e.g., in the absence of an adjuvant). In some embodiments, immunogenicity of a pharmaceutical composition is enhanced by including an adjuvant. Any adjuvant may be used in accordance with the present invention. A large number of adjuvants are known; a useful compendium of many such compounds is prepared by the National Institutes of Health and can be found (www.niaid.nih.gov/daids/vaccine/pdf/compendium.pdf). See also Allison, 1998, Dev. Biol. Stand., 92:3-11, Unkeless et al., 1998, Annu. Rev. Immunol., 6:251-281, and Phillips et al., 1992, Vaccine, 10:151-158. Hundreds of different adjuvants are known in the art and may be employed in the practice of the present invention. Exemplary adjuvants that can be utilized in accordance with the invention include, but are not limited to, cytokines, gel-type adjuvants (e.g., aluminum hydroxide, aluminum phosphate, calcium phosphate, etc.), microbial adjuvants (e.g., immunomodulatory DNA sequences that include CpG motifs; endotoxins such as monophosphoryl lipid A; exotoxins such as cholera toxin,  E. coli  heat labile toxin, and pertussis toxin; muramyl dipeptide, etc.), oil-emulsion and emulsifier-based adjuvants (e.g., Freund&#39;s Adjuvant, MF59 [Novartis], SAF, etc.), particulate adjuvants (e.g., liposomes, biodegradable microspheres, saponins, etc.), synthetic adjuvants (e.g., nonionic block copolymers, muramyl peptide analogues, polyphosphazene, synthetic polynucleotides, etc.) and/or combinations thereof. Other exemplary adjuvants include some polymers (e.g., polyphosphazenes; described in U.S. Pat. No. 5,500,161, Q57, Q S21, squalene, tetrachlorodecaoxide, etc. 
     In one embodiment, a pharmaceutical composition comprises a VLP of the invention formulated with aluminum phosphate adjuvant. Referring to Example 11, a triple modified SARS-CoV-2 spike protein (SEQ ID: 24) formulated with aluminum phosphate adjuvant (0.33 mg/ml) in a drug substance referred to as VBI-2902a was administered to human subjects in a Phase 1a clinical study. Interim data taken at day 56 showed that all subjects receiving two doses seroconverted after the second dose. 
     In some embodiments, pharmaceutical compositions are provided in a form that can be refrigerated and/or frozen. In some embodiments, pharmaceutical compositions are provided in a form that cannot be refrigerated and/or frozen. In some embodiments, reconstituted solutions and/or liquid dosage forms may be stored for a certain period of time after reconstitution (e.g., 2 hours, 12 hours, 24 hours, 2 days, 5 days, 7 days, 10 days, 2 weeks, a month, two months, or longer). In some embodiments, storage of VLP formulations for longer than the specified time results in VLP degradation. 
     A pharmaceutical composition in accordance with the invention may be prepared, packaged, and/or sold as a single unit dose, and/or as a plurality of single unit doses. As used herein, a “unit dose” is discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to a dose which would be administered to a subject and/or a convenient fraction of such a dose such as, for example, one-half or one-third of such a dose. 
     Relative amounts of active ingredient, pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition in accordance with the invention may vary, depending upon the identity, size, and/or condition of the subject and/or depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w/w) active ingredient. 
     Provided compositions and methods of the present disclosure are useful for prophylaxis and/or treatment of SARS, MERS or COVID-19 infection in a subject, including human adults and children. In general however they may be used with any animal. If desired, the methods herein may also be used with farm animals, such as ovine, avian, bovine, porcine and equine breeds. For the purposes of the present disclosure, vaccination can be administered before, during, and/or after exposure to a disease-causing agent, and in certain embodiments, before, during, and/or shortly after exposure to the agent. In some embodiments, vaccination includes multiple administrations, appropriately spaced in time, of a vaccinating composition. 
     Compositions described herein will generally be administered in such amounts and for such a time as is necessary or sufficient to induce an immune response. Dosing regimens may consist of a single dose or a plurality of doses over a period of time. The exact amount of an immunogenic composition (e.g., VLP) to be administered may vary from subject to subject and may depend on several factors. Thus, it will be appreciated that, in general, the precise dose used will depend not only on the weight of the subject and the route of administration, but also on the age of the subject. In certain embodiments a particular amount of a VLP composition is administered as a single dose. In certain embodiments, a particular amount of a VLP composition is administered as more than one dose (e.g., 1-3 doses that are separated by 1-12 months). 
     In some embodiments, a provided composition is administered in an initial dose and in at least one booster dose. In some embodiments, a provided composition is administered in an initial dose and two, three or four booster doses. In some embodiments, a provided composition is administered in an initial dose and in at least one booster dose about one month, about two months, about three months, about four months, about five months, or about six months following the initial dose. In some embodiments, a provided composition is administered in a second booster dose about six months, about seven months, about eight months, about nine months, about ten months, about eleven months, or about one year following the initial dose. In some embodiments, a provided composition is administered in a booster dose every 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years. 
     In a preferred embodiment, a composition comprising a VLP which expresses a triple modified SARS-CoV-2 spike protein based on the Beta variant (SEQ ID NO: 30) is used as a booster vaccine following a first vaccination with a composition comprising a triple modified SARS-CoV-2 spike protein based on the ancestral strain of SARS-CoV-2 (SEQ ID NO: 24). 
     In certain embodiments, provided compositions may be formulated for delivery parenterally, e.g., by injection. In such embodiments, administration may be, for example, intravenous, intramuscular, intradermal, or subcutaneous, or via by infusion or needleless injection techniques. In certain embodiments, the compositions may be formulated for peroral delivery, oral delivery, intranasal delivery, buccal delivery, sublingual delivery, transdermal delivery, transcutaneous delivery, intraperitoneal delivery, intravaginal delivery, rectal delivery or intracranial delivery. 
     In some embodiments, upon administration to a subject, provided VLPs induce a humoral immune response in the subject. In some embodiments, the humoral immune response in a subject is sustained for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 19 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 24 months, at least about 28 months, at least about 32 months, at least about 36 months, at least about 40 months, at least about 44 months, at least about 48 months, or longer. 
     In some embodiments, upon administration to a subject, provided VLPs induce a cellular immune response in the subject. In some embodiments, the cellular immune response in a subject is sustained for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least 12 months. 
     All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described herein. 
     EXAMPLES 
     The following examples describe some exemplary modes of making and practicing certain compositions that are described herein. It should be understood that these examples are for illustrative purposes only and are not meant to limit the scope of the compositions and methods described herein. 
     Example 1: Construction of DNA Expression Plasmids 
     This Example describes development of expression plasmids and constructs for expression of recombinant coronavirus spike gene sequences. A standard expression plasmid generally consists of a promoter sequence of mammalian origin, an intron sequence, a PolyAdenylation signal sequence (PolyA), a pUC origin of replication sequence (pUC—pBR322 is a colE1 origin/site of replication initiation and is used to replicate plasmid in bacteria such as  E. Coli  (DH5α)), and an antibiotic resistance gene as a selectable marker for plasmid plaque selection. Within the plasmid following the intron are a variety of restriction enzyme sites that can be used to splice in a gene or partial gene sequence of interest. 
     The Propol II expression plasmid contains the pHCMV (early promoter for HCMV), a Beta-Globin Intron (BGL Intron), a rabbit Globin polyAdenylation signal sequence (PolyA), a pUC origin of replication sequence (pUC—pBR322 is a colE1 origin/site of replication initiation and is used to replicate plasmid in bacteria such as  E. coli  (DH5α)), and an ampicillin resistance gene β-lactamase (Amp R—selectable marker for plasmid confers resistance to ampicillin (100 μg/ml). 
     To develop a Gag MMLV expression construct (“MLV-Gag”), a complementary DNA (cDNA) sequence encoding a Gag polyprotein of MMLV (Gag without its C terminus Pol sequence) (SEQ ID NO: 3) was cloned in a Propol II expression vector. To develop all of coronavirus expression constructs, each of the following sequences:
         i) SARS-CoV-2 (SEQ ID NO: 6);   ii) SARS-CoV (SEQ ID NO: 9);   iii) MERS (SEQ ID NO: 12);   iv) SARS-CoV-2 Proline Modified Spike Glycoprotein (SEQ ID NO: 15);   v) SARS-CoV-2 Furin Cleavage Modified (SEQ ID NO: 18);   vi) SARS-CoV-2 Proline and Furin Cleavage Modified (SEQ ID NO: 21);   vii) SARS-CoV-2 Proline and Furin Cleavage Modified with TM/Cyt from VSV (SEQ ID NO: 24); and   viii) SARS-CoV-2 modified with TM/Cyt from VSV (SEQ ID NO: 26)
 
was cloned in a Propol II expression vector. The SARS-CoV-2 sequence was from the L strain of the virus that was originally identified in Wuhan China.
       

     DNA plasmids were amplified in competent  E. coli  (DH5a) and purified with endotoxin-free preparation kits according to standard protocols. 
     Example 2: Production of Virus-Like Particles 
     This Example describes methods for production of virus-like particles containing various recombinant coronavirus spike antigens described in Example 1. 
     293 SF-3F6 cell line derived from HEK 293 cells are a proprietary suspension cell culture grown in serum-free chemically defined media (CA 2,252,972 and U.S. Pat. No. 6,210,922). HEK 293 SF-3F6 cells were scaled up in shaker flasks at 37° C., 5% CO 2  at a speed of 80 rpm and subsequently seeded in a bioreactor using HyQSF4 Transfx293 media supplemented with L-glutamine (GE Bioscience) to obtain a target cell density of 0.9 to 1.2 million cells/ml and high viability (&gt;90%). The cells were co-transfected at cell density of about ˜1 million cells/ml with different ratios of plasmids encoding coronavirus envelope polypeptides, plasmids encoding Gag and using high quality polyethyleneimine (PEIpro™) as transfection agent. The DNA plasmids and transfection agent were prepared in OptiPRO SFM medium (GE Biosciences). The bioreactor was monitored daily (˜24 hrs and 48 hrs post transfection) and cell density, viability and cell diameters recorded. The production broth was harvested at 48 hrs post transfection. 
     Total protein was determined on an aliquot by a Bradford assay quantification kit (BioRad). The Bradford Protein assay is based on the observation that the absorbance maximum for an acidic solution of Coomassie Brilliant Blue G-250 shifts from 465 nm to 595 nm when binding to protein occurs. Both hydrophobic and ionic interactions stabilize the anionic form of the dye, causing a visible color change. A spectrophotometer was used to measure the absorbance of the sample and Bradford Protein Reagent dye at 595 nm. 
     Example 3: Production of Monovalent Vaccine Candidates 
     Four different monovalent virus like particles were produced using the method described in Example 2. The virus like particles were transfected with one of the four following SARS-CoV-2 nucleotide sequences:
         1. Native form of SARS-CoV-2 (SEQ ID NO: 6);   2. SARS-CoV-2 Proline and Furin Cleavage Modified (SEQ ID NO: 21);   3. SARS-CoV-2 Proline and Furin Cleavage Modified with TM/Cyt from VSV (SEQ ID NO: 24); or   4. SARS-CoV-2 modified with TM/Cyt from VSV (SEQ ID NO: 26).       

     The total antigen content of the resulting products was measured and the results are shown in Table 1. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 Virus Like Particle Yields 
               
            
           
           
               
               
               
               
            
               
                   
                   
                   
                 SARS-CoV-2 
               
               
                   
                   
                 Gag 
                 spike protein 
               
               
                 Group 
                 Monovalent VLP Description 
                 (μg) 
                 (μg) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 1 
                 Native SARS-Cov-2 (SEQ ID #6) 
                 23,157 
                 16.2 
               
               
                 2 
                 SARS-CoV-2 Proline and Furin 
                 32,220 
                 229.9 
               
               
                   
                 Cleavage Modified (SEQ ID NO: 21) 
               
               
                 3 
                 SARS-CoV-2 Proline and Furin 
                 22,916 
                 639.2 
               
               
                   
                 Cleavage Modified with TM/Cyt 
               
               
                   
                 from VSV (SEQ ID NO: 24) 
               
               
                 4 
                 SARS-CoV-2 modified with TM/Cyt 
                 19,332 
                 49.95 
               
               
                   
                 from VSV (SEQ ID NO: 26) 
               
               
                   
               
            
           
         
       
     
     As can be seen from the data in Table 1, a significantly higher yield was obtained using Group 3, the SARS-CoV-2 sequence which had been modified by replacing the cytoplasmic and transmembrane segments with the corresponding segments form VSV. 
     Example 4: Production of Trivalent Vaccine Candidates 
     Four different trivalent virus like particles were produced using the method described in Example 2. Each particle was transfected with plasmids encoding Gag, an antigenic sequence from MERS (SEQ ID NO: 12), an antigenic sequence from SARS-CoV (SEQ ID NO: 9) and one of the two following SARS-CoV-2 sequences:
         1. Native form of SARS-CoV-2 envelope polypeptide (SEQ ID NO: 6); or   2. SARS-CoV-2 Proline and Furin Cleavage Modified with TM/Cyt from VSV (SEQ ID NO: 24).       

     The antigen content of the resulting products was measured and the results are shown in Table 2. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Trivalent Coronavirus Virus Like Particle Yields 
               
            
           
           
               
               
               
               
            
               
                   
                   
                   
                 SARS-CoV-2 
               
               
                   
                 Trivalent VLP - SARS- 
                 Gag 
                 spike protein 
               
               
                 Group 
                 CoV-2 Spike Protein 
                 (μg) 
                 (μg) 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 1 
                 Native SARS-Cov-2 
                 20,358 
                 28.8 
               
               
                   
                 (SEQ ID #6) 
               
               
                 2 
                 SARS-CoV-2 Proline 
                 17,382 
                 404.4 
               
               
                   
                 and Furin Cleavage 
               
               
                   
                 Modified with TM/Cyt 
               
               
                   
                 from VSV (SEQ ID 
               
               
                   
                 NO: 24) 
               
               
                   
               
            
           
         
       
     
     As can be seen from the data in Table 2, a significantly higher yield of trivalent VLPs was obtained using Group 2, the SARS-CoV-2 sequence with a stabilized prefusion form of the spike protein which was further modified with the TM/Cyt from VSV G protein (SEQ ID NO: 24). 
     Example 5: Evaluation of the Potency of Monovalent SARS-CoV-2 VLP Vaccine Constructs 
     Naïve 6-8 week-old C57/BL6 mice (n=10) were immunized twice with approximately 1/20 th  to 1/50 th  the human dose of the SARS-CoV-2 VLP vaccine formulations shown below in Table 3. Immunization took place at day 0 and day 21. Animals were sacrificed 14 days after immunization and their serum was collected for subsequent analysis of anti-spike protein antibody titers, and neutralizing antibodies. 
     The SARS-CoV-2 VLPs were formulated with aluminum phosphate adjuvant (Adjuphos®) as shown in Table 3. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 VLP Vaccine Formulations 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Dose 
               
               
                   
                   
                 μg SARS- 
                   
                   
                 volume/ 
               
               
                   
                   
                 CoV-2 
                 μg 
                 Al+++ 
                 Animal 
               
               
                 Group 
                 Monovalent SARS-CoV-2 
                 Spike/dose 
                 Gag/dose 
                 μg/dose 
                 (μL) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 1 
                 SARS-CoV-2 native Spike protein VLP 
                 0.07 
                 100.1 
                 125 
                 500 
               
               
                 2 
                 SARS-CoV-2 Proline and Furin 
                 0.14 
                 19.6 
                 125 
                 500 
               
               
                 3 
                 Cleavage Modified (SEQ ID NO: 21) 
                 0.7 
                 98.1 
                 125 
                 500 
               
               
                 4 
                 SARS-CoV-2 Proline and Furin 
                 0.14 
                 5.0 
                 125 
                 500 
               
               
                 5 
                 Cleavage Modified with TM/Cyt 
                 0.7 
                 25.05 
                 125 
                 500 
               
               
                   
                 from VSV (SEQ ID NO: 24) 
               
               
                 6 
                 SARS-CoV-2 modified with 
                 0.07 
                 27.1 
                 125 
                 500 
               
               
                 7 
                 TM/Cyt from VSV (SEQ ID NO: 26) 
                 0.14 
                 54.2 
                 125 
                 500 
               
               
                   
               
            
           
         
       
     
     Anti-Spike SARS-CoV-2 antibody titers were measured as follows: 96 well plates were coated overnight at 4° C., with SARS-COV-2 Spike Protein (51+S2) (Sinobiological, Cat #40589-V08B1) (0.1 μg/ml in DPBS). The following day, plates were blocked with 5% milk in ELISA wash buffer, for 1 hour at 37° C. Plates were washed with wash buffer, followed by addition of 2 fold dilutions of individual mouse sera starting at 1:10,000 to 1:1,200,000. Plates were incubated for 1.5 hours at 37° C., followed by plate washing and addition of Secondary Antibody: Goat anti-Mouse IgG1 (Bethyl, Cat #A90-131P), diluted 1:5,000 in 1% milk in ELISA wash buffer. Plates were incubated for 1 hour at 37° C. Plates were added with TMB One component Microwell substrate, incubated at room temperature for 10 minutes and then added with Stop solution. Absorbance was read at 450 nm using a MAXline plate reader. Results are shown below in Table 4. 
     The anti-spike total IgG binding titers reported in Table 4 represent the highest dilution of sera that still had an optical density of 0.1 or greater by ELISA measurement against recombinant SARS-CoV-2 spike protein. Unexpectedly, immunization of mice with just a single dose of VLPs expressing the stabilized prefusion form of the SARS-CoV-2 spike protein further modified with the TM/Cyt from the VSV-G protein (Group 4) induced antibody responses which were dramatically stronger than immunization of mice with VLPs expressing similar doses of SARS-CoV-2 spike protein but in different presentations (Groups 2, 7). 
     The antibody titers from the mice 14 days after each vaccination are shown in Table 4. P1 and P2 refer to the first and second vaccination. Results were pooled among individual animals. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 VLP Vaccine Antibody Titres 
               
            
           
           
               
               
               
               
            
               
                   
                   
                   
                 Anti- SARS-CoV-2 Spike 
               
               
                   
                 Group 
                 Vaccination 
                 Total IgG Binding Titers 
               
               
                   
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Pooled Group 1 
                 P1Vd14 
                 9,099 
               
               
                   
                   
                 P2Vd14 
                 310,103 
               
               
                   
                 Pooled Group 2 
                 P1Vd14 
                 74,612 
               
               
                   
                   
                 P2Vd14 
                 424,883 
               
               
                   
                 Pooled Group 3 
                 P1Vd14 
                 262,689 
               
               
                   
                   
                 P2Vd14 
                 321,427 
               
               
                   
                 Pooled Group 4 
                 P1Vd14 
                 341,493 
               
               
                   
                   
                 P2Vd14 
                 670,735 
               
               
                   
                 Pooled Group 5 
                 P1Vd14 
                 619,766 
               
               
                   
                   
                 P2Vd14 
                 359,528 
               
               
                   
                 Pooled Group 6 
                 P1Vd14 
                 1,108 
               
               
                   
                   
                 P2Vd14 
                 302,500 
               
               
                   
                 Pooled Group 7 
                 P1Vd14 
                 4,093 
               
               
                   
                   
                 P2Vd14 
                 221,990 
               
               
                   
                   
               
            
           
         
       
     
     As is shown in Table 4, each of the monovalent VLP vaccine formulations induced a strong antibody response in mice. In almost all formulations, the response was strongly enhanced by a second vaccination. One group, group 5 consisting of a vaccine formulation based on SARS-CoV-2 Proline and Furin Cleavage Modified with TM/Cyt from VSV (SEQ ID NO: 24), showed a reduced response after second vaccination. However, the response was very high after first vaccination, raising the possibility that the second vaccination exhausted the immune response in mice. It is possible that this response may not be seen in larger mammals such as humans. 
     Neutralizing antibodies were tested as follows. A constant amount of virus consisting of 100 plaque forming units (pfu) of a Canadian isolate of SARS-CoV-2 virus was mixed with 2-fold dilutions of the mouse serum specimens being tested, the dilutions ranging from 40 to 5120 times, followed by plating of the mixture onto cells of an appropriate cell line for the individual virus. The concentration of plaque forming units is determined by the number of plaques formed after a few days. A vital dye (e.g. crystal violet or neutral red) was then added for visualization of the plaques and the number of plaques in an individual plate with test serum was divided by the number of plaques present in a negative control sera to calculate the percentage neutralization. The plaque forming units were measured by microscopic observations or by observation of specific dyes that react with the infected cell. Interpretation is typically based on 50% neutralization, which is the last dilution of serum capable of inhibiting 50% of the total plaques (virions). Plaque reduction neutralization test (PRNT) thresholds of 80 and 90 represent dilutions of sera capable of reducing plaques by 80% or 90% respectively. The results are shown in Table 5. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 VLP Vaccine Neutralizing Antibodies 
               
            
           
           
               
               
               
               
            
               
                 Test Group 
                 PRNT50 
                 PRNT80 
                 PRNT90 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 POV (before vaccination) 
                 ** 
                   
                   
               
               
                 P1VD14 Group 1 
                 ** 
               
               
                 P2VD14 Group 1 
                 2560 
                 1280 
                 640 
               
               
                 P1VD14 Group 2 
                  320 
                 160 
                 80 
               
               
                 P2VD14 Group 2 
                 *** 
                 5120 
                 2560 
               
               
                 P1VD14 Group 3 
                  320 
                 285 
                 160 
               
               
                 P2VD14 Group 3 
                 5120 
                 2560 
                 1280 
               
               
                 P1VD14 Group 4 
                  640 
                 320 
                 160 
               
               
                 P2VD14 Group 4 
                 *** 
                 5120 
                 2560 
               
               
                 P1VD14 Group 5 
                 2560 
                 640 
                 320 
               
               
                 P2VD14 Group 5 
                 5120 
                 2276 
                 1280 
               
               
                 P1VD14 Group 6 
                 ** 
               
               
                 P2VD14 Group 6 
                 5120 
                 2560 
                 640 
               
               
                 P1VD14 Group 7 
                  160 
                 80 
                 ** 
               
               
                 P2VD14 Group 7 
                 2560 
                 1280 
                 320 
               
               
                   
               
               
                 ** below lowest dilution limit of PRNT (titer 40); 
               
               
                 *** above highest dilution range of PRNT (titer 5120) 
               
            
           
         
       
     
     As shown in Table 5, all of the monovalent vaccine constructs induced a neutralizing antibody response. This response was very potent, as demonstrated by the data from the stringent PRNT 90 threshold. 
     Example 6: Evaluation of the Potency of a Trivalent SARS-CoV-2 VLP Vaccine Construct 
     A trivalent VLP was prepared using the method in Example 2 with antigen plasmids including all of the following sequences: 
     
       
         
           
               
               
            
               
                   
                 i) 
               
               
                   
                 (SEQ ID NO: 6) 
               
               
                   
                 SARS-CoV-2; 
               
               
                   
                   
               
               
                   
                 ii) 
               
               
                   
                 (SEQ ID NO: 9) 
               
               
                   
                 SARS-CoV; 
               
               
                   
                 and 
               
               
                   
                   
               
               
                   
                 iii) 
               
               
                   
                 (SEQ ID NO: 12) 
               
               
                   
                 MERS. 
               
            
           
         
       
     
     Vaccine formulations comprising the trivalent VLP, a monovalent VLP (expressing native SARS-CoV-2 (SEQ ID NO. 6), a recombinant SARS-CoV-2 (SEQ ID NO: 25) and Gag protein alone (SEQ ID NO:1) were tested in vivo in mice. The recombinant SARS-CoV-2 (SEQ ID NO: 25) was provided by the National Research Council of Canada. The vaccines were formulated with aluminum phosphate adjuvant (Adjuphos®) as shown in Table 6. 
     Forty naïve 6-8 week-old C57/BL6 mice (4 groups of 10) were immunized three times with approximately 1/20 th  to 1/50 th  of a human dose of the vaccine formulations shown below in Table 6. Immunization took place at day 0, day 21 and day 42. Animals were sacrificed 14 days after the last immunization and their serum was collected for subsequent analysis of anti-spike protein antibody titers and neutralizing antibodies. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Vaccine Formulations 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Dose 
               
               
                   
                   
                 μg SARS- 
                   
                   
                 volume/ 
               
               
                 Test 
                   
                 CoV-2 
                 μg 
                 Al+++ 
                 Animal 
               
               
                 Group 
                 Description 
                 Spike/dose 
                 Gag/dose 
                 pg/dose 
                 (μL) 
               
               
                   
               
               
                 1 
                 Monovalent SARS-CoV-2 native spike 
                 0.1 
                 50 
                 125 
                 250 
               
               
                   
                 protein VLP 
               
               
                 2 
                 Trivalent SARS-CoV-2 (native spike 
                 0.1 
                 50 
                 125 
                 250 
               
               
                   
                 protein); SARS-CoV; MERS-CoV VLP 
               
               
                 3 
                 SARS-CoV-2 spike recombinant 
                 0.1 
                 N/A 
                 125 
                 250 
               
               
                 4 
                 Empty Gag 
                 N/A 
                 50 
                 125 
                 250 
               
               
                   
               
            
           
         
       
     
     Anti-Spike SARS-CoV-2, anti-SARS and anti-MERS antibody titers were measured for each group using the technique described in Example 5 with the following capture antigens (SARS-COV-2 Spike Protein (51+S2), Sino Biological, Cat #40589-V08B1, SARS-COVSpike Protein (51+S2), MyBioSource, Cat #MBS434077 and MERS-CoV Spike Protein (51+S2), Sino Biological, Cat #40069-V08B). The results are shown in Table 7. 
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Coronavirus Antibody Titres 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Anti SARS- 
                 Anti SARS- 
                 Anti MERS 
               
               
                   
                 Time 
                 CoV2 Spike 
                 CoV Spike 
                 Spike 
               
               
                 Group 
                 Point 
                 binding titre 
                 binding titre 
                 binding titre 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 Pooled Group 1 
                 P1Vd14 
                 2700 
                 300 
                 negative 
               
               
                 Monovalent 
                 P2Vd14 
                 72900 
                 8100 
                 100 
               
               
                 VLP 
                 P3Vd14 
                 218700 
                 24300 
                 negative 
               
               
                 Pooled Group 2 
                 P1Vd14 
                 2700 
                 900 
                 &gt;2700 
               
               
                 Trivalent 
                 P2Vd14 
                 24300 
                 24300 
                 &gt;72900 
               
               
                 VLP 
                 P3Vd14 
                 &gt;72900 (72900~ 
                 72900 
                 656100 
               
               
                   
                   
                 218700) 
               
               
                 Pooled Group 3 
                 P1Vd14 
                 2700 
                 900 
                 negative 
               
               
                 Stabilized 
                 P2Vd14 
                 72900 
                 24300 
                 900 
               
               
                 Recombinant 
                 P3Vd14 
                 218700 
                 24300 
                 negative 
               
               
                 SARS-CoV2 
               
               
                 Pooled Group 4 
                 P1Vd14 
                 negative 
                 negative 
                 negative 
               
               
                 Empty Gag 
                 P2Vd14 
                 negative 
                 negative 
                 negative 
               
               
                 VLP 
                 P3Vd14 
                 negative 
                 negative 
                 negative 
               
               
                   
               
            
           
         
       
     
     As shown in Table 7, the trivalent VLP (Group 2) induced antibody responses against all three coronaviruses: SARS-CoV-2, SARS and MERS. This demonstrates that a trivalent vaccine candidate has the potential to provide immunological protection again all three major coronaviruses. 
     Anti-SARS-CoV-2 binding and PRNT 80 neutralizing titres for individual animals after the third vaccination are shown in Table 8 below. Neutralizing antibodies were measured using the method described in Example 5. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Anti-SARS-CoV-2 Binding and Neutralizing Titres for Individual Mice 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Anti 
                   
                   
                   
               
               
                   
                   
                   
                   
                 SARS-CoV2 
                 Anti 
               
               
                   
                   
                   
                 Anti 
                 binding 
                 SARS-CoV2 
                 Neutralizing 
                 nAb/ 
               
               
                   
                   
                   
                 SARS-CoV2 
                 titre 
                 Neutralizing 
                 Antibodies 
                 Binding 
               
               
                 Group 
                   
                 Mouse 
                 binding 
                 Geometric 
                 Antibodies 
                 Geometric 
                 Titres 
               
               
                 # 
                 Description 
                 # 
                 titre 
                 Mean 
                 (PRNT 80) 
                 Mean 
                 Ratio 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 1 
                 SARS-CoV-2 
                 6 
                 1,057,003 
                 355538 
                 2560 
                 1810 
                 0.002 
               
               
                   
                 native Spike 
                 7 
                 221,691 
                   
                 640 
                   
                 0.003 
               
               
                   
                 protein eVLP 
                 8 
                 515,381 
                   
                 1280 
                   
                 0.002 
               
               
                   
                   
                 9 
                 240,199 
                   
                 5120 
                   
                 0.021 
               
               
                   
                   
                 10 
                 124,759 
                   
                 1280 
                   
                 0.01 
               
               
                   
                   
                 11 
                 1,320,203 
                   
                 5120 
                   
                 0.004 
               
               
                   
                   
                 12 
                 258,417 
                   
                 1280 
                   
                 0.005 
               
               
                   
                   
                 13 
                 252,174 
                   
                 640 
                   
                 0.003 
               
               
                   
                   
                 14 
                 454,574 
                   
                 5120 
                   
                 0.011 
               
               
                   
                   
                 15 
                 228,032 
                   
                 1280 
                   
                 0.006 
               
               
                 2 
                 SARS-CoV-2 + 
                 21 
                 333,263 
                 247080 
                 &gt;5120 
                 &gt;5120 
                 0.015 
               
               
                   
                 SARS-CoV- + 
                 22 
                 299,575 
                   
                 &gt;5120 
                   
                 0.017 
               
               
                   
                 MERS-CoV 
                 23 
                 208,191 
                   
                 &gt;5120 
                   
                 0.024 
               
               
                   
                 Native Spike 
                 24 
                 142,458 
                   
                 &gt;5120 
                   
                 0.036 
               
               
                   
                 protein eVLP 
                 25 
                 294,434 
                   
                 &gt;5120 
                   
                 0.017 
               
               
                   
                   
                 26 
                 129,136 
                   
                 &gt;5120 
                   
                 0.04 
               
               
                   
                   
                 27 
                 322,558 
                   
                 &gt;5120 
                   
                 0.016 
               
               
                   
                   
                 28 
                 390,847 
                   
                 &gt;5120 
                   
                 0.013 
               
               
                   
                   
                 29 
                 278,663 
                   
                 &gt;5120 
                   
                 0.019 
               
               
                   
                   
                 30 
                 214,394 
                   
                 &gt;5120 
                   
                 0.024 
               
               
                   
               
            
           
         
       
     
     As can be seen from the data shown in Table 8 demonstrates that the trivalent VLP induced higher neutralizing antibody responses than the monovalent SARS-CoV-2 VLP even though the binding titres were lower. This is particularly evident when by observing the ratio f neutralizing antibodies to binding antibody titres in the last column of Table 8. This demonstrates that the trivalent vaccine candidate has the potential to provide stronger immunological protection against COVID-19. 
     The serum obtained from mice fourteen (14) days after each vaccination was tested for cross reactivity with a different coronavirus which is known to infect humans and cause a common cold (HCoV-0C43). Antibody titres were measured using ELISA as described above using human coronavirus (HCoV-0C43) spike protein (S1+S2 ECD, His Tag), Sino, Cat #40607-V08B, stock 0.25 mg/mL as the capture antigen. The results are shown below in Table 9 below. 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Cross Reactivity of Mouse Serum 
               
               
                 against HCoV-OC43 Spike Protein 
               
            
           
           
               
               
               
               
            
               
                   
                   
                   
                 Anti-HCoV-OC43 
               
               
                   
                   
                 Vaccination 
                 Spike Total IgG 
               
               
                   
                 Group 
                 Number 
                 Binding Titres 
               
               
                   
                   
               
               
                   
                 Pooled Group 1 
                 P1VD14 
                 negative 
               
               
                   
                 Monovalent VLPs 
                 P2VD14 
                 100 
               
               
                   
                   
                 P3VD14 
                 300 
               
               
                   
                 Pooled Group 2 
                 P1VD14 
                 negative 
               
               
                   
                 Trivalent VLPs 
                 P2Vd14 
                 900 
               
               
                   
                   
                 P3VD14 
                 2700  
               
               
                   
                 Pooled Group 3 NRC 
                 P1VD14 
                 negative 
               
               
                   
                 Stabilized Recombinant 
                 P2VD14 
                 300 
               
               
                   
                 SARS-CoV2 
                 P3Vd14 
                 300 
               
               
                   
                 Pooled Group 4 Empty 
                 P1VD14 
                 negative 
               
               
                   
                 eVLP 
                 P2VD14 
                 negative 
               
               
                   
                   
                 P3VD14 
                 negative 
               
               
                   
                   
               
            
           
         
       
     
     As can be seen from Table 9 above, the trivalent VLP vaccine candidate (Group 2) demonstrated higher cross reactivity against a human coronavirus which causes common cold. As such, the trivalent candidate demonstrated the potential for broader protection against coronavirus than the monovalent VLP or the recombinant SARS-CoV-2 spike protein alone. 
     In order to evaluate the efficacy of the vaccine formulations, the neutralizing antibodies were also measured in human serum (HS) collected from four recovered COVID-19 patients and the results were compared to the neutralizing antibodies induced by the four different test groups shown in Table 6. PRNT 50 and PRNT 90 was determined following the first and second vaccination using the method described in Example 5. Pooled results for each group are shown in Table 10 below. 
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Anti-SARS-CoV-2 Neutralizing Antibodies 
               
            
           
           
               
               
               
               
            
               
                   
                 Sample 
                 PRNT50 
                 PRNT90 
               
               
                   
                   
               
               
                   
                 HS1 
                  80 
                 ** 
               
               
                   
                 HS2 
                 160 
                 40 
               
               
                   
                 HS3 
                 1280  
                 320  
               
               
                   
                 HS4 
                 160 
                 80 
               
               
                   
                 POV (BEFORE VACCINATION) 
                 ** 
                 ** 
               
               
                   
                 P1VD14 GR1 
                 ** 
                 ** 
               
               
                   
                 P1VD14 GR2 
                 ** 
                 ** 
               
               
                   
                 P1VD14 GR3 
                  40 
                 ** 
               
               
                   
                 P1VD14 GR4 
                 ** 
                 ** 
               
               
                   
                 P2VD14 GR1 
                 640 
                 160  
               
               
                   
                 P2VD14 GR2 
                 320 
                 ** 
               
               
                   
                 P2VD14 GR3 
                 640 
                 80 
               
               
                   
                 P2VD14 GR4 
                 ** 
                 ** 
               
               
                   
                   
               
               
                   
                 ** below lowest dilution limit of PRNT (titer 40); 
               
            
           
         
       
     
     As can be seen in Table 10, the monovalent VLP vaccine induced more neutralizing antibodies than COVID-19 infection in three out of four human patients as measured by PRNT 50 and 90. The trivalent VLP vaccine induced more neutralizing antibodies than COVID-19 infection in three out of four human patients as measured by PRNT 50. Accordingly, the vaccine constructs at least as effective, and potentially more effective, at inducing immune protection than exposure to SARS-CoV-2. 
     Example 7: Evaluation of Protective Effect of a Monovalent SARS-CoV-2 VLP Vaccine Construct 
     Syrian golden hamsters (males, aged approximately 5-6 weeks old) were divided into two groups and immunized with two doses of the formulations shown below in Table 11, specifically a test sample comprising a triple modified SARS-CoV2 VLP (SEQ ID: 24) formulated with aluminum phosphate adjuvant (Adjuphos®) (Group B) and a saline control (Group A). Immunizations took place at day 0 and day 21, via intramuscular injection. At day 42, all animals were challenged intranasally with 50 μl of SARS-CoV-2 via both nares, at a challenge virus dose of 1×10 5  TCID 50  per animal. SST (serum separation tube) blood samples (approximately 0.5 ml each) were collected on day 0 prior to the prime immunization, day 14 and day 35, respectively. Final blood samples were collected at necropsy. Nasal washes were collected on days 35, 43, 44, 45, 47, 49, 51, 53 and 56. Half of the animals in each group were euthanized at three days post-challenge, and the remaining animals were euthanized at 14 days post-challenge. 
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 VLP Vaccine Formulations 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Dose 
               
               
                   
                   
                 μg SARS- 
                   
                 volume/ 
               
               
                   
                   
                 CoV-2 
                 Al+++ 
                 Animal 
               
               
                 Group 
                 Test Article 
                 Spike/dose 
                 μg/dose 
                 (μL) 
               
               
                   
               
               
                 A 
                 Saline control 
                 N/A 
                 N/A 
                 100 
               
               
                 B 
                 SARS-CoV-2 Proline 
                 1 
                 125 
                 100 
               
               
                   
                 and Furin Cleavage 
               
               
                   
                 Modified with TM/Cyt 
               
               
                   
                 from VSV (SEQ ID 
               
               
                   
                 NO: 24) 
               
               
                   
               
            
           
         
       
     
     At necropsy, gross lung pathology was evaluated and the proportion of lung lobe that contained lesions was estimated. Lung tissues were analyzed for viral load by qRT-PCR and viral cell culture. Similarly, nasal turbinates were collected for viral load by qRT-PCR and viral cell culture. 
     Extraction of RNA from nasal washes was performed using Qiagen reagents (QIAamp Viral RNA Mini Kit Cat No./ID: 52906). Briefly, 140 μl of nasal wash was added into 560 μl viral lysis buffer (Buffer AVL). The mixture was incubated at room temperature for 10 min. After brief centrifugation, the solution was transferred to a fresh tube containing 560 μL of 100% ethanol, and the tube was incubated at room temperature for 10 min. RNA was then purified and eluted with 60 μl of RNase Free water containing 0.04% sodium azide (elution buffer AVE). 
     Extraction of RNA from lung lobes and nasal turbinates was completed using approximately 100 μg of tissue. The tissues were homogenized in 600 μl of lysis buffer (RLT Qiagen) with a sterile stainless steel bead in the TissueLyserII (Qiagen) for 6 min, at 30 Hz. The solution was centrifuged at 5000×g for 5 min. Supernatant was transferred to a fresh tube containing 600 μl of 70% ethanol, and the tube was incubated at room temperature for 10 min. Viral RNA was then purified using Qiagen RNeasy Mini Kit (Cat No/ID: 74106) and eluted with 50 μL elution buffer. 
     qRT-PCR assays were performed on RNA from samples of nasal washes, lung tissues and nasal turbinates using SARS-CoV-2 specific primers (Table 12). The primers had an annealing temperature of approximately 60° C. Qiagen Quantifast RT-PCR Probe kits were used for qRT-PCR, and the qRT-PCR reactions were conducted using the OneStep Plus (Applied Biosystems) machine. The qRT-PCR results were expressed in copy number per reaction, by producing a standard curve with a sample of a linearized plasmid DNA that contains the env gene of SARS-CoV-2. The Ct values for individual samples were used with the standard curve to determine the copy number in each sample. 
     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 Sequence of Primers Used 
               
            
           
           
               
               
               
            
               
                 Primer 
                 Sequence 
                 SEQ ID NO: 
               
               
                   
               
               
                 Forward Primer (Fwd) 
                 ACAGGTACGTTAATAGTTAATAGCGT 
                 31 
               
               
                   
               
               
                 Reverse Primer (Rev) 
                 ATATTGCAGCAGTACGCACACA 
                 32 
               
               
                   
               
               
                 Labelled Probe 
                 ACACTAGCCATCCTTACTGCGCTTCG 
                 33 
               
               
                   
               
            
           
         
       
     
     Viral titration assays were performed to assess infectious virus. The assays were conducted in 96-well plates using Vero′76 cells (ATCC CRL-1587). Median tissue culture infectious dose (TCID 50 ) was determined by microscopic observation of the cytopathic effect (CPE) of cells. The virus was quantified and reported in TCID 50 /ml or TCID 50 /gram. TCID 50  values were calculated using the Spearman &amp; Karber algorithm in Excel. 
     Anti-Spike SARS-CoV-2 antibody titers were measured by ELISA performed on serum samples. Plates were coated with spike S1+S2 Ag (Cat #40589-V08B1, Sino Biological Inc.). The coating concentration was 0.1 ug/mL. Plates were blocked with 5% non-fat skim milk powder in PBS containing 0.05% Tween 20. Fourfold dilutions of serum were used. Goat anti-Hamster IgG HRP from ThermoFisher (PA1-29626) was used as the secondary antibody at 1:7000. Plates were developed with OPD peroxidase substrate (0.5 mg/ml) (Thermo Scientific Pierce 34006). The reaction was stopped with 2.5 M sulfuric acid and absorbance was measured at 490 nm. Throughout the assay, plates were washed with PBS containing 0.05% Tween 20. The assay was performed in duplicate. The titres were reported as the end point of the dilutions. 
     Antibodies to the spike protein receptor binding domain (“RBD”) were measured as follows. Anti-SARS-CoV-2 spike 51 RBD IgG antibody binding titer was determined from serum samples using an indirect ELISA. Recombinant SARS-CoV-2 spike 51 RBD protein was adsorbed on a microtiter plate overnight and plates were then blocked with a solution of 5% skim milk in wash buffer for 1 hour. After blocking and washing, samples were added to the microplates and incubated for 1.5 hours. An HRP-conjugated goat anti-Syrian Hamster IgG-Fc was used as a detection antibody, and incubated on the microplates for 1 hour. The signal was developed with Tetramethylbenzidine (TMB) substrate solution and the reaction stopped by addition of 450 μL Liquid Stop Solution for TMB Microwell Substrate. The absorbance was read at 450 nm using an ELISA microwell plate reader. 
     Viral neutralization assays against the challenge SARS-CoV-2 virus were performed on the serum samples using the cell line Vero′76. The serum samples were heat-inactivated for 30 min at 56° C. The serum samples were serially diluted (2-fold serial dilutions). The experiment was conducted in technical duplicates. The virus was diluted in medium to a concentration of 25 TCID 50  in 50 μl per well (the inoculum size=25 TCID 50 ). Then 60 μl of the virus solution was mixed with 60 μl serially diluted serum samples. The mixture was incubated for 1 hr at 37° C., with 5% CO 2 . The pre-incubated virus-serum mixtures (100 μl) were transferred to the wells of the 96-well flat-bottom plates containing 90% confluent pre-seeded Vero′76 cells. The plates were incubated at 37° C., with 5% CO 2  for five days. The plates were observed using a microscope on day 1 post-infection (dpi) for contamination and on days 3 and 5 post-infection for cytopathic effect. The serum dilution factor for the wells with no CPE at 5 dpi was defined as the serum neutralization titre. The initial serum dilution factor was 1:20. 
     Neutralizing antibodies were tested as follows. Vero cells were seeded at 8×10 5  cells/well in 6-well plates 48 h prior to infection. Sera were heat-inactivated at 56° C. for 30 min then transferred on ice. Sera were diluted 1:10 with virus infection media then each diluted serum was used to carry out ½× fold serial dilutions to give 1:20 to 1:40960 (8 subsequent dilutions). Equal volumes of diluted serum and virus (100 pfu per serum dilution) were mixed and incubated at 37° C. for 1 h. No sera and no virus controls were included. Cells were washed with PBS and each virus/serum were transferred and mixed to each well containing cells, and incubated at 37° C. for 1 h, with interval rocking of the plates. After the 1 h adsorption, excess inoculum was removed and a 2 ml virus infection media/agarose mix were overlaid onto the cells. The overlay was allowed to solidify and plates were incubated at 37° C. for 72 h. Cells were stained with crystal violet at 72 h post-infection. Plaques were quantified for all dilutions and PRNT titer was calculated. The % plaque reduction for all the dilutions based on the no serum control, was calculate using the Reed-Muench formula to determine the PRNT titers 50, 80, and 90. 
     Lung tissues were also quantified for cytokine gene expression collected at necropsy. The gene expression of IL-4, IL-10, IL-13, TNF-alpha and IFN-gamma was determined in the right cranial and right caudal lung lobe by qRT-PCR using the primers shown in Table 13. The beta-actin gene expression was used for reference. 
     
       
         
           
               
             
               
                 TABLE 13 
               
             
            
               
                   
               
               
                 Primer Sequence 
               
            
           
           
               
               
               
               
               
            
               
                 Gene 
                   
                 SEQ 
                   
                 SEQ 
               
               
                 Target 
                 Forward Primer 5′→3′ 
                 ID NO: 
                 Reverse Primer 5′→3′ 
                 ID NO: 
               
               
                   
               
               
                 IL-4 
                 CCACGGAGAAAGACCTCA 
                 34 
                 GGGTCACCTCATGTTGGAAA 
                 40 
               
               
                   
                 TCTG 
                   
                 TAAA 
                   
               
               
                   
               
               
                 IL-10 
                 GTTGCCAAACCTTATCAGA 
                 35 
                 TTCTGGCCCGTGGTTCTCT 
                 41 
               
               
                   
                 AATGA 
                   
                   
                   
               
               
                   
               
               
                 IL-13 
                 AAATGGCGGGTTCTGTGC 
                 36 
                 AATATCCTCTGGGTCTTGTAG 
                 42 
               
               
                   
                   
                   
                 ATGG 
                   
               
               
                   
               
               
                 TNF- 
                 GGAGTGGCTGAGCCATCGT 
                 37 
                 AGCTGGTTGTCTTTGAGAGA43 
                   
               
               
                 alpha 
                   
                   
                 CATG 
                   
               
               
                   
               
               
                 IFN- 
                 GGCCATCCAGAGGAGCAT 
                 38 
                 TTTCTCCATGCTGCTGTTGAA 
                 44 
               
               
                 gamma 
                 AG 
                   
                   
                   
               
               
                   
               
               
                 Beta- 
                   
                   
                   
                   
               
               
                 actin 
                 ACTGCCGCATCCTCTTCCT 
                 39 
                 TCGTTGCCAATGGTGATGAC 
                 45 
               
               
                   
               
            
           
         
       
     
     Lung tissues were collected in RNAlater and the RNA was isolated using Qiagen RNeasy Mini extraction kits using RLT lysis buffer (Qiagen RNeasy Mini Kit, Cat No/ID:74106). RNA concentration and the 260/280 ratio as an indicator of purity was determined by a nanodrop spectrophotometer. cDNA was synthesized using iScript™ Reverse Transcription Supermix with 500 ng of RNA as template. cDNA was synthesized following a program of 5 min at 25° C., 20 min at 46° C., and 95° C. for 1 min. Master mix was prepared for each gene of interest as well as a house keeping gene at 10% overage: 1.84 μl Nuclease Free H 2 O; Forward Primer 0.08 μL; Reverse Primer 0.08 μl; and SYBR 10 μl [SYBR® Green PCR Master Mix (SsoAdvanced™ Universal SYBR® Green Supermix #1725275)]. Twelve μl of the master mix was combined with 8 μl of RNA for each PCR reaction. After loading, the plate was centrifuged at 1500 RPM for 1 min to bring all liquid back into base of well. The qPCR was performed using a Bio-Rad Thermocycler (Bio-Rad CX1000). Data was analyzed using the Bio-Rad CFX Maestro software. Data is exported in the form of Ct values to an excel spreadsheet for fold change calculation by ΔΔCt Formula in Excel. 
     Results based on clinical observation of animals indicated that all animals were healthy throughout the immunization phase. All animals had normal activity levels and had no clinical signs. The body weight increases were normal in the group vaccinated with test article (Group B) when compared to the Saline control group (Group A). 
     Immune response to vaccination as measured by antibody titres to SARS-CoV-2 spike protein are shown in Table 14 fourteen days after the first vaccination and fourteen days after the second vaccination (P1 and P2 refer to the first and second vaccination). Results shown are Geo means of the animals in each group. 
     
       
         
           
               
             
               
                 TABLE 14 
               
             
            
               
                   
               
               
                 SARS-CoV-2 VLP Vaccine Antibody Titres 
               
            
           
           
               
               
               
            
               
                   
                 First 
                 Second 
               
               
                   
                 Vaccination (day 14) 
                 Vaccination (day 14) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Anti- 
                 Anti- 
                 Anti- 
                 Anti- 
               
               
                   
                 SARS- 
                 SARS- 
                 SARS- 
                 SARS- 
               
               
                   
                 CoV-2 
                 CoV-2 
                 CoV-2 
                 CoV-2 
               
               
                   
                 S1 
                 Spike 
                 S1 
                 Spike 
               
               
                   
                 RBD 
                 total IgG 
                 RBD 
                 Total IgG 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                 Group A 
                 — 
                 3 
                 — 
                 3 
               
               
                 (control) 
               
               
                 Group B 
                 7,446 
                 1,222 
                 268,399 
                 22,868 
               
               
                 (SARS-CoV-2 
               
               
                 Proline and 
               
               
                 Furin Cleavage 
               
               
                 Modified with 
               
               
                 TM/Cyt from VSV 
               
               
                 (SEQ ID NO: 24) 
               
               
                   
               
            
           
         
       
     
     The Group B animals (immunized with the triple modified monovalent SARS-CoV-2 VLP vaccine) produced high levels of anti-spike antibody two weeks after the second vaccination. At two weeks after the first vaccination, 10 out of 12 animals in Group B produced anti-spike antibodies (data not shown). Group A animals (Saline control) did not have anti-spike antibody production. The triple modified monovalent SARS-CoV-2 VLP vaccine also induced detectable level of anti-SARS-CoV-2-S1 RBD IgG antibody at 14 days after the first immunization. A substantial increase in antibody titres was observed on day 14 after the 2 nd  immunization. No anti-SARS-CoV-2-S1 RBD IgG were detected in control Group A. 
     The neutralizing antibodies, as determined by PRNT, for each group fourteen days after the first vaccination are shown in Table 15 (average values shown). Values indicate reciprocal of highest dilution that showed inhibition of 50% (PRNT50), 80% (PRNT80), or 90% (PRNT90) of input virus, respectively. 
     
       
         
           
               
             
               
                 TABLE 15 
               
             
            
               
                   
               
               
                 SARS-CoV-2 VLP Vaccine Neutralizing Antibodies 
               
            
           
           
               
               
               
               
            
               
                 Test Group - Fourteen days 
                   
                   
                   
               
               
                 after first vaccination 
                 PRNT50 
                 PRNT80 
                 PRNT90 
               
               
                   
               
               
                 Group A Pooled 
                 ** 
                 (N/A) 
                 (N/A) 
               
               
                 (Control) 
               
               
                 Group B 
                 441 
                 241 
                 198 
               
               
                 (SARS-CoV-2 Proline and 
               
               
                 Furin Cleavage Modified 
               
               
                 with TM/Cyt from VSV 
               
               
                 (SEQ ID NO: 24) 
               
               
                   
               
               
                 ** below lowest dilution limit of PRNT (titer 40) 
               
            
           
         
       
     
     All Group B animals produced virus neutralizing antibodies at two weeks post-immunization as shown in Table 15. The Group A animals did not produce any neutralizing antibodies as shown in Table 15. 
     At three days post-challenge (dpc), all animals Group B, and only one animal in Group A, produced neutralizing antibodies (data not shown). At 14 dpc, all the animals in Group A and B produced neutralizing antibodies. (data not shown). 
     During the challenge phase, all animals except for two were active and had normal activity levels, and did not have abnormal nasal signs. 
     Animals were weighed each day post challenge. After challenge, Group A animals lost approximately 15% of their initial body weight, peaking at 6-8 dpc. The means of % body weight changes of the Group B animals were only about 1-2% and peaked at two dpc. Body weight data is shown in Table 16 below at Day 0 and at Day 3 and 6 after challenge. 
     
       
         
           
               
             
               
                 TABLE 16 
               
             
            
               
                   
               
               
                 Body Weight of Hamsters following Viral Challenge 
               
            
           
           
               
               
               
               
            
               
                   
                 Day 0 
                 Day 3 
                 Day 6 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                 Animal 
                   
                 BW 
                   
                 BW 
                   
                 BW 
               
               
                 Group 
                 ID 
                 BW 
                 Average 
                 BW 
                 Average 
                 BW 
                 Average 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 A 
                 401 
                 175.1 
                 171.5 ± 14.5 
                 160.9 
                 160.9 ± 13.9 
                 Euth 
                 155.5 ± 11.5 
               
               
                 (control) 
                 403 
                 182.1 
                   
                 167.3 
                   
                 155.4 
               
               
                   
                 408 
                 172.6 
                   
                 160.7 
                   
                 Euth 
               
               
                   
                 412 
                 173.6 
                   
                 161 
                   
                 149.1 
               
               
                   
                  416* 
                 165.9 
                   
                 163.2 
                   
                 137.1 
               
               
                   
                 422 
                 196.3 
                   
                 185.8 
                   
                 170.5 
               
               
                   
                 423 
                 162.8 
                   
                 149.6 
                   
                 Euth 
               
               
                   
                 429 
                 185.2 
                   
                 171.6 
                   
                 159.6 
               
               
                   
                 434 
                 174.8 
                   
                 170.00 
                   
                 161.5 
               
               
                   
                 435 
                 156.4 
                   
                 143.7 
                   
                 Euth 
               
               
                   
                 440 
                 174.3 
                   
                 164.9 
                   
                 Euth 
               
               
                   
                 443 
                 139.4 
                   
                 132.1 
                   
                 Euth 
               
               
                 B 
                 402 
                 159.5 
                 174.2 ± 14.3 
                 155.7 
                 171.6 ± 14.3 
                 Euth 
                 180.3 ± 7.9  
               
               
                 (SARS-CoV-2 
                 404 
                 181.2 
                   
                 177.1 
                   
                 179.6 
               
               
                 Proline and 
                 410 
                 161.4 
                   
                 160.1 
                   
                 Euth 
               
               
                 Furin Cleavage 
                 411 
                 190.0 
                   
                 183.6 
                   
                 182.1 
               
               
                 Modified with 
                 418 
                 181.3 
                   
                 179.1 
                   
                 185.1 
               
               
                 TM/Cyt from 
                  421* 
                 188.4 
                   
                 189.5 
                   
                 191.6 
               
               
                 VSV (SEQ ID 
                 426 
                 160.0 
                   
                 159.0 
                   
                 Euth 
               
               
                 NO: 24) 
                 428 
                 175.8 
                   
                 173.3 
                   
                 172.8 
               
               
                   
                 431 
                 172.3 
                   
                 169.1 
                   
                 170.3 
               
               
                   
                 436 
                 152.7 
                   
                 147.3 
                   
                 Euth 
               
               
                   
                 442 
                 199.1 
                   
                 195.4 
                   
                 Euth 
               
               
                   
                 446 
                 168.8 
                   
                 169.9 
                   
                 Euth 
               
               
                   
               
               
                 Euthanized on day 3 as planned or humane euthanized 
               
               
                 *(416 and 421) 
               
            
           
         
       
     
     As can be seen in Table 16, animals given the Saline solution lost considerable weight three days and six days after challenge whereas the animals that had received the vaccine lost considerably less weight at day 3 and were had gained weight by day 6. 
     Viral RNA as measured in nasal washes post challenge is shown in Table 17. In all days examined, the vaccinated (Group B) animals had lower viral RNA levels in nasal washes than the Group A animals (control group), as depicted in Table 17 (showing copies/Rxn for each day post-challenge). Only during day two after challenge were the viral RNA levels significantly lower in Groups B compared to Group A (p=0.0206). 
     
       
         
           
               
             
               
                 TABLE 17 
               
             
            
               
                   
               
               
                 Viral RNA in nasal washes 
               
            
           
           
               
               
            
               
                   
                 Table 16 Day post-challenge Viral RNA Averages (copies/Rxn) 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Group ID 
                 0 
                 2 
                 3 
                 5 
                 7 
                 9 
                 11 
                 14 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Group A 
                 0.1 
                 3708602 
                 340897 
                 94932 
                 12409 
                 6231 
                 130 
                 3146 
               
               
                 Group B 
                 0.1 
                 205616 
                 105469 
                 31861 
                 93 
                 20 
                 2461 
                 2 
               
               
                   
               
            
           
         
       
     
     Viral RNA at 3 days post-challenge in various tissues for control (Group A) and vaccinated (Group B) animals are shown in Table 18 (showing values for copies/gram). At three days post-challenge, viral RNA was detectable in the right cranial lobe (RCra) and the right caudal lobe (RCau) of the lung and the nasal turbinates in all animals. When compared to Group A, the levels of viral RNA in the RCra of Group B were significantly lower. Similarly, the levels of RNA in RCau were significantly lower in Group B than Group A. In the nasal turbinates, viral RNA levels in Group B was significantly lower than in Group A. 
     
       
         
           
               
             
               
                 TABLE 18 
               
             
            
               
                   
               
               
                 Viral RNA in tissues at 3 days post-challenge 
               
            
           
           
               
               
               
               
            
               
                   
                 Right cranial 
                 Right caudal 
                 Nasal 
               
               
                 Group ID 
                 lobe 
                 lobe 
                 turbinates 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Group A 
                 2613650533 
                 1608083300 
                 2099767910 
               
               
                 (control) 
               
               
                 Group B 
                 108006 
                 42465 
                 1205108163 
               
               
                 (SARS-CoV-2 Proline and 
               
               
                 Furin Cleavage Modified 
               
               
                 with TM/Cyt from VSV 
               
               
                 (SEQ ID NO: 24) 
               
               
                   
               
            
           
         
       
     
     Viral RNA at 14 days post-challenge in various tissues for the control (Group A) and vaccinated (Group B) animals are shown in Table 19 (showing values for copies/gram). At 14 days post-challenge, viral RNA was detectable in all Group A animals and some animals in Group B in the RCra, RCau or nasal turbinates. The levels of RNA in RCra and RCau were significantly different in Group B than those in Group A. 
     
       
         
           
               
             
               
                 TABLE 19 
               
             
            
               
                   
               
               
                 Viral RNA in Tissues at 14 days Post-challenge 
               
            
           
           
               
               
               
               
            
               
                   
                 Right cranial 
                 Right caudal 
                 Nasal 
               
               
                 Group ID 
                 lobe 
                 lobe 
                 turbinates 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Group A 
                 127855.5 
                 62075.0 
                 3104415.2 
               
               
                 (control) 
               
               
                 Group B 
                 99.43 
                 105.78 
                 100398.57 
               
               
                 (SARS-CoV-2 Proline and 
               
               
                 Furin Cleavage Modified 
               
               
                 with TM/Cyt from VSV 
               
               
                 (SEQ ID NO: 24) 
               
               
                   
               
            
           
         
       
     
     Infectious virus in various tissues at 3 days post-challenge for control (Group A) and vaccinated (Group B) animals are shown in Table 20 (showing values for TCID 50 /gram). At three days post-challenge, infectious virus was detectable in all animals of Group A in the right cranial and right caudal lobes of the lung and in nasal turbinates. The titres of infectious virus in Group B was significantly lower than those in Group A. At 14 days post-challenge, infectious virus was not detected in any of the animals (data not shown). 
     
       
         
           
               
             
               
                 TABLE 20 
               
             
            
               
                   
               
               
                 Infectious Virus in tissues at 3 days post-challenge (TCID50/gram) 
               
            
           
           
               
               
               
               
            
               
                   
                 Right cranial lobe 
                 Right caudal lobe 
                 Nasal turbinates 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Group ID 
                 TCID50/gram 
                 Average 
                 TCID50/gram 
                 Average 
                 TCID50/gram 
                 Average 
               
               
                   
               
               
                 Group A 
                 1.51E+07 
                 2.63E+07 
                 6.71E+02 
                 1.65E+06 
                 1.15E+07 
                 2.00E+07 
               
               
                 (control) 
                 1.03E+07 
                   
                 3.94E+05 
                   
                 2.39E+06 
               
               
                   
                 3.89E+07 
                   
                 3.45E+05 
                   
                 3.76E+07 
               
               
                   
                 1.30E+07 
                   
                 1.23E+06 
                   
                 4.66E+05 
               
               
                   
                 1.32E+06 
                   
                 5.99E+06 
                   
                 2.47E+07 
               
               
                 Group B 
                 1.79E+01 
                 1.48E+01 
                 8.78E+00 
                 3.45E+00 
                 1.26E+02 
                 3.58E+06 
               
               
                 (SARS-CoV-2 
                 1.49E+01 
                   
                 2.56E+00 
                   
                 5.43E+05 
               
               
                 Proline and 
                 2.39E+01 
                   
                 2.28E+00 
                   
                 5.86E+01 
               
               
                 Furin Cleavage 
                 9.05E+00 
                   
                 2.42E+00 
                   
                 2.09E+07 
               
               
                 Modified with 
                 1.12E+01 
                   
                 2.59E+00 
                   
                 6.85E+00 
               
               
                 TM/Cyt from 
               
               
                 VSV (SEQ ID 
               
               
                 NO: 24) 
               
               
                   
               
            
           
         
       
     
     Heavier lungs is associated with more advanced disease. Therefore, the ratio of lung weight to body is correlated with more severe disease states. Table 21 shows the lung weight to body weight ratios for animals in the control (Group A) and vaccinated (Group B) animals three days post challenge. Animals in group Group B animals had significantly lower lung weight to body weight ratios. 
     
       
         
           
               
             
               
                 TABLE 21 
               
             
            
               
                   
               
               
                 Ratios of lung weight to body weight (%) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Lung 
                 Body 
                 Lung weight/ 
                   
               
               
                   
                 Weight 
                 Weight 
                 Body Weight 
                 Average ± 
               
               
                 Group ID 
                 (g) 
                 (g) 
                 Ratio 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 Group A 
                 1.4 
                 160.9 
                 0.87 
                 0.76 ± 0.15 
               
               
                 (control) 
                 1.2 
                 170.1 
                 0.71 
               
               
                   
                 1.5 
                 160.7 
                 0.93 
               
               
                   
                 1.1 
                 160.9 
                 0.68 
               
               
                   
                 0.88 
                 137.1 
                 0.64 
               
               
                   
                 1.4 
                 183.7 
                 0.76 
               
               
                   
                 1.6 
                 149.6 
                 1.07 
               
               
                   
                 1.3 
                 183.6 
                 0.71 
               
               
                   
                 1 
                 182.3 
                 0.55 
               
               
                   
                 1.1 
                 143.7 
                 0.77 
               
               
                   
                 1.3 
                 164.9 
                 0.79 
               
               
                   
                 0.8 
                 132.1 
                 0.61 
               
               
                 Group B 
                 0.9 
                 155.7 
                 0.58 
                 0.56 ± 0.07 
               
               
                 (SARS-CoV-2 
                 1 
                 188.2 
                 0.53 
               
               
                 Proline and Furin 
                 1 
                 160.1 
                 0.62 
               
               
                 Cleavage Modified 
                 0.9 
                 188.2 
                 0.48 
               
               
                 with TM/Cyt from 
                 0.9 
                 195.2 
                 0.46 
               
               
                 VSV (SEQ ID 
                 0.98 
                 191.6 
                 0.51 
               
               
                 NO: 24) 
                 1 
                 159 
                 0.63 
               
               
                   
                 0.9 
                 180.1 
                 0.50 
               
               
                   
                 1 
                 183.6 
                 0.54 
               
               
                   
                 1 
                 147.3 
                 0.68 
               
               
                   
                 1.2 
                 195.4 
                 0.61 
               
               
                   
                 NA 
                 169.9 
                 missing 
               
               
                   
               
            
           
         
       
     
     Following necropsy, lung tissues were fixed in formalin, embedded, sectioned and stained with hematoxylin and eosin (H&amp;E). Slides were examined by a board-certified pathologist and scored on a scale of 0-4 as shown in Table 22. 
     
       
         
           
               
             
               
                 TABLE 22 
               
             
            
               
                   
               
               
                 Lung Histology Scores (median) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 Intensity of 
                 Extent of 
                   
                   
               
               
                   
                   
                 Proportion of 
                 the inflammatory 
                 hypertrophy 
               
               
                 Group 
                 Days Post- 
                 parenchyma 
                 infiltrate in 
                 of alveolar 
                 Interalveolar 
                 Extent of 
               
               
                 ID 
                 Challenge 
                 affected 
                 affected areas 
                 pneumocytes 
                 hemorrhage 
                 emphysema 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Group A 
                 3 
                 3 
                 3 
                 0 
                 2 
                 0 
               
               
                 (control) 
                   
                 3 
                 3 
                 0 
                 1 
                 0 
               
               
                   
                   
                 3 
                 3 
                 0 
                 2 
                 0 
               
               
                   
                   
                 1 
                 3 
                 0 
                 3 
                 0 
               
               
                   
                   
                 1 
                 3 
                 0 
                 2 
                 0 
               
               
                   
                   
                 1 
                 2 
                 0 
                 1 
                 0 
               
               
                   
                 14 
                 1 
                 2 
                 3 
                 0 
                 0 
               
               
                   
                   
                 1 
                 2 
                 2 
                 0 
                 0 
               
               
                   
                   
                 4 
                 4 
                 3 
                 4 
                 0 
               
               
                   
                   
                 4 
                 2 
                 2 
                 0 
                 0 
               
               
                   
                   
                 2 
                 3 
                 2 
                 0 
                 0 
               
               
                   
                   
                 1 
                 2 
                 1 
                 0 
                 0 
               
               
                 Group B 
                 3 
                 0 
                 0 
                 0 
                 0 
                 1 
               
               
                 (SARS-CoV-2 
                   
                 1 
                 2 
                 0 
                 0 
                 0 
               
               
                 Proline and 
                   
                 1 
                 1 
                 0 
                 0 
                 0 
               
               
                 Furin Cleavage 
                   
                 1 
                 2 
                 0 
                 0 
                 0 
               
               
                 Modified with 
                   
                 1 
                 1 
                 0 
                 1 
                 0 
               
               
                 TM/Cyt from 
                   
                 1 
                 1 
                 0 
                 0 
                 0 
               
               
                 VSV (SEQ ID 
                 14 
                 0 
                 0 
                 0 
                 0 
                 0 
               
               
                 NO: 24) 
                   
                 0 
                 0 
                 0 
                 0 
                 0 
               
               
                   
                   
                 0 
                 0 
                 0 
                 0 
                 0 
               
               
                   
                   
                 1 
                 1 
                 0 
                 0 
                 0 
               
               
                   
                   
                 0 
                 0 
                 0 
                 0 
                 0 
               
               
                   
                   
                 0 
                 0 
                 0 
                 0 
                 0 
               
               
                   
               
            
           
         
       
     
     As can be seen in Table 22, animals in the control group (Group A) showed significant disease pathology following challenge at days 3 and 14. By way of contrast, the vaccinated animals (Group B) showed some minor pathology at day 3 but were mostly recovered by day 14. Accordingly, the vaccine provided significant protection against disease induced lung pathology. 
     Immunohistochemical staining was conducted to observe SARS-CoV-2 virus in the lung tissues, specifically the parenchyma and bronchioles/bronchi. Staining was observed and scores for the two groups of animals is shown in Table 23. 
     
       
         
           
               
             
               
                 TABLE 23 
               
             
            
               
                   
               
               
                 Immunohistochemical scores (median) 
               
               
                 of the lung for SARS-CoV-2 
               
            
           
           
               
               
               
            
               
                   
                 Parenchyma IHC 
                 Bronchioles/bronchi 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Indi- 
                   
                 Indi- 
                   
               
               
                 Group 
                 Days post- 
                 vidual 
                   
                 vidual 
               
               
                 ID 
                 challenge 
                 Score 
                 Median 
                 Score 
                 Median 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 3 
                 4 
                   4 ± 0.84 
                 2 
                 3 ± 0.52 
               
               
                 (Control) 
                   
                 4 
                   
                 3 
               
               
                   
                   
                 4 
                   
                 2 
               
               
                   
                   
                 4 
                   
                 3 
               
               
                   
                   
                 3 
                   
                 3 
               
               
                   
                   
                 2 
                   
                 3 
               
               
                   
                 14 
                 0 
                     0±1.21 
                 2 
                 2±0.75 
               
               
                   
                   
                 0 
                   
                 2 
               
               
                   
                   
                 3 
                   
                 1 
               
               
                   
                   
                 1 
                   
                 1 
               
               
                   
                   
                 0 
                   
                 3 
               
               
                   
                   
                 0 
                   
                 2 
               
               
                 Group B 
                 3 
                 0 
                 0 ± 0 
                 2 
                 1 ± 0.89 
               
               
                 (SARS-CoV-2 
                   
                 0 
                   
                 1 
               
               
                 Proline and 
                   
                 0 
                   
                 2 
               
               
                 Furin Cleavage 
                   
                 0 
                   
                 0 
               
               
                 Modified with 
                   
                 0 
                   
                 1 
               
               
                 TM/Cyt from 
                   
                 0 
                   
                 0 
               
               
                 VSV (SEQ ID 
                 14 
                 0 
                 0 ± 0 
                 0 
                 1 ± 0.63 
               
               
                 NO: 24) 
                   
                 0 
                   
                 1 
               
               
                   
                   
                 0 
                   
                 2 
               
               
                   
                   
                 0 
                   
                 1 
               
               
                   
                   
                 0 
                   
                 1 
               
               
                   
                   
                 0 
                   
                 1 
               
               
                   
               
            
           
         
       
     
     Vaccinated animals had significantly less virus stain in both parenchyma and bronchioles/bronchi than those of the saline control animals (Group A). At 14 days post-challenge, virus stain was similar among the groups in either parenchyma or bronchioles/bronchi although still a little lower in the vaccinated group. 
     The transcriptional levels of cytokines IL-4, IL-10, IL-13, TNF-alpha and IFN-gamma in the right cranial lung, right caudal lung and the nasal turbinates were determined by qRT-PCR. At 3 days post-challenge, IL-10, IL-13 and IFN-gamma displayed differential expression in the right cranial lobe and the right caudal lobe in Group B (shown in Tables 24 and 25). In nasal turbinates, IL-10 and IFN-gamma exhibited differential expression (shown in Table 26). At 14 days post-challenge, the transcriptional levels of IL-4, IL-10, IL-13, TNF-alpha and IFN-gamma in the right cranial lung, right caudal lung and the nasal turbinates were similar across the groups (shown in Tables 27-29). 
     
       
         
           
               
             
               
                 TABLE 24 
               
             
            
               
                   
               
               
                 Transcriptional profiles of cytokines in right cranial 
               
               
                 lobe 3 days post-challenge (fold changes) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group ID 
                 IL-4 
                 IL-10 
                 IL-13 
                 TNF-alpha 
                 IFN-gamma 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 1.04 
                 1.09 
                 1.3 
                 1.07 
                 1.14 
               
               
                 Group B 
                 1.8 
                 0.37 
                 7.1 
                 0.61 
                 0.27 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 25 
               
             
            
               
                   
               
               
                 Transcriptional profiles of cytokines in right caudal 
               
               
                 lobe 3 days post-challenge (fold changes) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group ID 
                 IL-4 
                 IL-10 
                 IL-13 
                 TNF-alpha 
                 IFN-gamma 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 1.02 
                 1.03 
                 1.33 
                 1.04 
                 1.21 
               
               
                 Group B 
                 0.88 
                 0.33 
                 2.4 
                 0.55 
                 0.35 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 26 
               
             
            
               
                   
               
               
                 Transcriptional profiles of cytokines in nasal 
               
               
                 turbinates 3 days post-challenge (fold changes) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group ID 
                 IL-4 
                 IL-10 
                 IL-13 
                 TNF-alpha 
                 IFN-gamma 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 1.22 
                 1.05 
                 1.26 
                 1.16 
                 1.09 
               
               
                 Group B 
                 1.44 
                 0.84 
                 2.87 
                 0.45 
                 0.82 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 27 
               
             
            
               
                   
               
               
                 Transcriptional profiles of cytokines in right cranial 
               
               
                 lobe 14 days post-challenge (fold changes) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group ID 
                 IL-4 
                 IL-10 
                 IL-13 
                 TNF-alpha 
                 IFN-gamma 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 1.07 
                 1.25 
                 1.57 
                 1.34 
                 1.24 
               
               
                 Group B 
                 1.37 
                 0.92 
                 2.22 
                 0.77 
                 0.61 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 28 
               
             
            
               
                   
               
               
                 Transcriptional profiles of cytokines in right caudal 
               
               
                 lobe 14 days post-challenge (fold changes) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group ID 
                 IL-4 
                 IL-10 
                 IL-13 
                 TNF-alpha 
                 IFN-gamma 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 1.09 
                 1.51 
                 1.13 
                 1.11 
                 1.31 
               
               
                 Group B 
                 1.38 
                 0.78 
                 0.63 
                 0.94 
                 0.65 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 29 
               
             
            
               
                   
               
               
                 Transcriptional profiles of cytokines in nasal turbinates 
               
               
                 14 days post-challenge (fold changes) 
               
            
           
           
               
               
               
               
               
               
            
               
                 Group ID 
                 IL-4 
                 IL-10 
                 IL-13 
                 TNF-alpha 
                 IFN-gamma 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Group A 
                 1.46 
                 1.45 
                 1.43 
                 1.41 
                 1.46 
               
               
                 Group B 
                 0.73 
                 0.72 
                 1.40 
                 0.65 
                 0.57 
               
               
                   
               
            
           
         
       
     
     Example 8: Evaluation of Potency and Protective Effect of Single Dose of Monovalent SARS-CoV-2 VLP Vaccine Construct 
     Syrian golden hamsters (males, aged approximately 6-7 weeks old) were immunized with the monovalent triple modified SARS-CoV-2 VLP vaccine formulations shown below in Table 30. Immunizations took place only at day 21 via intramuscular injection. Serum was collected at day 0 and day 35 for subsequent analysis of neutralizing antibodies. 
     
       
         
           
               
             
               
                 TABLE 30 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 VLP Vaccine Formulations 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 μg 
                   
                 Dose 
               
               
                   
                   
                 SARS- 
                   
                 volume/ 
               
               
                   
                   
                 CoV-2 
                 Al+++ 
                 Animal 
               
               
                 Group 
                 Test Article 
                 Spike/dose 
                 μg/dose 
                 (μL) 
               
               
                   
               
               
                 A 
                 Saline control 
                 N/A 
                 N/A 
                 100 
               
               
                 B 
                 SARS-CoV-2 Proline and 
                 1.4 
                 125 
                 100 
               
               
                   
                 Furin Cleavage Modified 
               
               
                   
                 with TM/Cyt from VSV 
               
               
                   
                 (SEQ ID NO: 24) 
               
               
                   
               
            
           
         
       
     
     Neutralizing antibodies were tested using the plaque reduction neutralization test (PRNT), as described in Example 7 for animals in Group B. The results are shown in Table 31 (average values shown). 
     
       
         
           
               
             
               
                 TABLE 31 
               
             
            
               
                   
               
               
                 Monovalent SARS-CoV-2 VLP Vaccine Neutralizing Antibodies 
               
            
           
           
               
               
               
               
            
               
                 Test Group 
                 PRNT50 
                 PRNT80 
                 PRNT90 
               
               
                   
               
               
                 Day 0 (before vaccination) 
                 ** 
                 — 
                 — 
               
               
                 Day 14 after vaccination 
                 190 
                 65 
                 45 
               
               
                 Group B 
               
               
                   
               
               
                 ** below lowest dilution limit of PRNT (titer 40) 
               
            
           
         
       
     
     Compared to Group B of Example 7 (where animals received immunizations of 1 μg SARS-CoV-2 Spike/dose at day 0 and day 21), animals in Group B of this Example 8 (where animals received a single immunization of 1.4 μg SARS-CoV-2 Spike/dose at day 21) exhibited a higher serum neutralizing antibody response. These data support effective immunization with only a single dose of monovalent SARS-CoV-2 VLP vaccine. 
     Challenge studies were performed on day 42, as described in Example 7. Table 32 shows average body weights (grams) of animals before challenge. 
     
       
         
           
               
             
               
                 TABLE 32 
               
             
            
               
                   
               
               
                 Pre-Challenge Average Body Weights of Animals 
               
            
           
           
               
               
               
            
               
                 Day 
                 Group A 
                 Group B 
               
               
                   
               
            
           
           
               
               
               
            
               
                 −1 
                 113.8 
                 116.6 
               
               
                 7 
                 131.7 
                 134.6 
               
               
                 10 
                 136.3 
                 139.9 
               
               
                 22 
                 154.5 
                 157.6 
               
               
                 28 
                 160.4 
                 164.0 
               
               
                 35 
                 168.4 
                 172.0 
               
               
                 42 
                 174 
                 178.5 
               
               
                   
               
            
           
         
       
     
     Table 33 shows average body weights (grams) of animals post-challenge. As can be seen in Table 33, animals who received a single dose of vaccine lost less weight than those who received saline. 
     
       
         
           
               
             
               
                 TABLE 33 
               
             
            
               
                   
               
               
                 Post-Challenge Average Body Weights of Animals 
               
            
           
           
               
               
               
            
               
                 Day 
                 Group A 
                 Group B 
               
               
                   
               
            
           
           
               
               
               
            
               
                 1 
                 172.1 
                 174.6 
               
               
                 2 
                 168.3 
                 172.6 
               
               
                 3 
                 165.3 
                 173.4 
               
               
                 4 
                 163.7 
                 168.5 
               
               
                 5 
                 159.1 
                 168.5 
               
               
                 6 
                 155.7 
                 168.5 
               
               
                 7 
                 154.7 
                 170.1 
               
               
                 8 
                 157.5 
                 171.7 
               
               
                 9 
                 162.9 
                 173.8 
               
               
                 10 
                 164.4 
                 174.0 
               
               
                 11 
                 166.6 
                 175.3 
               
               
                 12 
                 169.4 
                 177.1 
               
               
                 13 
                 172.0 
                 177.8 
               
               
                 14 
                 173.5 
                 178.9 
               
               
                   
               
            
           
         
       
     
     Table 34 shows average % change in body weights of animals post-challenge. 
     
       
         
           
               
             
               
                 TABLE 34 
               
             
            
               
                   
               
               
                 Post-Challenge Average % Body Weight Change of Animals 
               
            
           
           
               
               
               
            
               
                 Day 
                 Group A 
                 Group B 
               
               
                   
               
            
           
           
               
               
               
            
               
                 1 
                 −1.09 
                 −2.24 
               
               
                 2 
                 −3.29 
                 −3.30 
               
               
                 3 
                 −5.08 
                 −2.90 
               
               
                 4 
                 −6.51 
                 −2.10 
               
               
                 5 
                 −9.14 
                 −2.15 
               
               
                 6 
                 −11.11 
                 −2.11 
               
               
                 7 
                 −11.64 
                 −1.22 
               
               
                 8 
                 −10.05 
                 −0.31 
               
               
                 9 
                 −7.03 
                 0.87 
               
               
                 10 
                 −6.22 
                 0.93 
               
               
                 11 
                 −4.94 
                 1.63 
               
               
                 12 
                 −3.35 
                 2.69 
               
               
                 13 
                 −1.89 
                 3.09 
               
               
                 14 
                 −1.00 
                 3.74 
               
               
                   
               
            
           
         
       
     
     These data demonstrate that a single immunization of 1.4 μg SARS-CoV-2 Spike/dose at day 21 was effective at preventing reduction in body weight following viral challenge, relative to control. 
     Example 9: Evaluation of Monovalent and Trivalent SARS-CoV-2 VLP Vaccine Constructs for Antibody Titers Against South African SARS-CoV-2 Variant 
     Monovalent and trivalent SARS-CoV-2 VLP vaccine constructs which have the triple modified SARS-CoV-2 spike protein were assessed for production of antibodies against South African SARS-CoV-2 variant. Mice were immunized IP twice (on day 0 and day 21, as described in Example 6) with the SARS-CoV-2 VLP vaccine formulations shown below in Table 35. Animals were sacrificed 14 days after immunization and their serum was collected for subsequent analysis of anti-spike protein antibody titers. 
     The SARS-CoV-2 VLPs were formulated with aluminum phosphate adjuvant (Adjuphos®) as shown in Table 35. 
     
       
         
           
               
             
               
                 TABLE 35 
               
             
            
               
                   
               
               
                 SARS-CoV-2 VLP Vaccine Formulations 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 μg 
                   
                 Dose 
               
               
                   
                   
                 SARS- 
                   
                 volume/ 
               
               
                   
                   
                 CoV-2 
                 Al+++ 
                 Animal 
               
               
                 Group 
                 Test Article 
                 Spike/dose 
                 μg/dose 
                 (μL) 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 1 
                 Monovalent VLP - SARS- 
                 0.2 
                 125 
                 500 
               
               
                   
                 CoV-2 Proline and Furin 
               
               
                   
                 Cleavage Modified with 
               
               
                   
                 TM/Cyt from VSV (SEQ 
               
               
                   
                 ID NO: 24) 
               
               
                 2 
                 Trivalent VLP (v4) - 
                 0.2 
                 125 
                 500 
               
               
                   
                 SARS-CoV-2 Spike Protein 
               
               
                   
                 (described in Example 4) 
               
               
                   
               
            
           
         
       
     
     Antibody titers were assessed by ELISA, as described in Example 7, except that well plates were coated with SARS-COV-2 Spike Protein from South African variant. Antibody titers at 14 days after the second immunizations are shown in Table 36. Results shown are Geo means of the animals in each group. 
     
       
         
           
               
             
               
                 TABLE 36 
               
             
            
               
                   
               
               
                 SARS-CoV-2 VLP Vaccine Antibody Titres 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Anti- SARS- 
               
               
                   
                   
                   
                 Anti- SARS- 
                 CoV-2 African 
               
               
                   
                   
                 Days post 
                 CoV-2 Spike 
                 Variant Spike 
               
               
                   
                   
                 Second 
                 Total IgG 
                 Total IgG 
               
               
                   
                 Group 
                 Vaccination 
                 Binding Titers 
                 Binding Titers 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Group 1 
                 14 
                 128,850 
                 121,511 
               
               
                   
                 Group 2 
                 14 
                 215,232 
                 211,080 
               
               
                   
                   
               
            
           
         
       
     
     These data demonstrate that mice injected with the monovalent and trivalent vaccines produced antibodies which bind to the South African variant of the Spike protein of SARS-CoV. 
     Example 10: Evaluation of Isotype Antibody Titer of Monovalent SARS-CoV-2 VLP Vaccine Construct 
     In another study, the isotype of antibody titers were assessed, following immunization of mice with the vaccine constructs shown in Table 37. 
     
       
         
           
               
             
               
                 TABLE 37 
               
             
            
               
                   
               
               
                 SARS-CoV-2 VLP Vaccine Antibody Titres 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 μg 
                   
                 Dose 
               
               
                   
                   
                 SARS- 
                   
                 volume/ 
               
               
                   
                   
                 CoV-2 
                 Al+++ 
                 Animal 
               
               
                 Group 
                   
                 Spike/dose 
                 μg/dose 
                 (μL) 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 1 
                 Monovalent Native 
                 0.2 
                 125 
                 500 
               
               
                   
                 SARS-Cov-2 (SEQ ID 
               
               
                   
                 NO: 6) VLP 
               
               
                 2 
                 Stabilized Recombinant 
                 0.2 
                 125 
                 500 
               
               
                   
                 SARS-CoV2 spike 
               
               
                   
                 protein (non-VLP) 
               
               
                   
               
            
           
         
       
     
     Mice were immunized IP twice (on day 0 and day 21, as described in Example 6). Animals were sacrificed 14 days after immunization and their serum was collected for subsequent analysis of anti-spike protein antibody titers. 
     As shown in Table 38, unexpectedly, when VLPs were formulated with the same amount/concentration of alum as recombinant spike protein, a balanced antibody response was seen (IgG1/IgG2b). Increased production of IgG2b is associated with a TH1 immune response, which is indicative of cell-mediated immunity. This indicates that the VLP construct resulted in elevated levels of IgG2b expression which is correlated to the more effective TH1 immune response. 
     
       
         
           
               
             
               
                 TABLE 38 
               
             
            
               
                   
               
               
                 SARS-CoV-2 VLP Vaccine Antibody Titres 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Anti- SARS- 
                 Anti- SARS- 
               
               
                   
                   
                 Days post 
                 CoV-2 Spike 
                 CoV-2 Spike 
               
               
                   
                   
                 second 
                 IgG1 
                 IgG2b 
               
               
                   
                 Group 
                 vaccination 
                 Binding Titers 
                 Binding Titers 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Group 1 
                 14 
                 172,105 
                 116,633 
               
               
                   
                 Group 2 
                 14 
                 198,469 
                 9,674 
               
               
                   
                   
               
            
           
         
       
     
     Example 11: A Phase 1a Randomized, Placebo Controlled Study to Evaluate the Monovalent SARS-CoV-2 VLP Vaccine in Human Subjects 
     An investigational vaccine called VBI-2902a was formulated using 5 μg of the SARS-CoV-2 Proline and Furin Cleavage Modified with TM/Cyt from VSV (SEQ ID NO: 24) spike protein with 0.33 mg/ml aluminum phosphate adjuvant (Adjuphos®). Sixty (60) healthy adults, 18-54 years of age with no previous clinical or laboratory diagnosis of COVID-19 or SARS-CoV-2 infection and not previously vaccinated with an experimental or authorized COVID-19 vaccine were enrolled in Phase 1a clinical study to evaluate the safety, tolerability and immunogenicity of a one-dose or two-dose regimen. Participants were randomized into the following groups:
         Group G1—20 participants received VBI-2902a at a dose of 5 μg of spike protein at Day 1 and placebo at Day 28;   Group G2—20 participants received VBI-2902a at a dose of 5 μg of spike protein at       

     Days 1 and 28; and
         Group G3—20 participants received placebo at Days 1 and 28.       

     All subjects were seronegative at the start of the trial. Immunogenicity (antibody binding and neutralization titers) was measured on Days 0, 7, 28, 35, and 56. Prior to Day 56 there were 11 individuals who chose to be vaccinated with COVID-19 vaccines available under emergency use authorization and there were 3 confirmed cases of SARS-CoV-2 infection (no infections occurred in the two-dose cohort). Immunogenicity results excluded these 14 subjects. 
     There were no antibody or neutralization responses detected in any subjects in the placebo group at any time point, except for a single subject at Days 35 and 56 which likely represented an undiagnosed case of COVID-19 given the relatively high antibody binding and neutralization titers in this subject. 
     Immunogenicity assessments utilized validated assays that assessed binding titers against a recombinant SARS-CoV-2 spike protein and neutralization titers based on a pseudovirus neutralization assay, with both assays using a Wuhan SARS-CoV-2 sequence. A panel of 25 COVID-19 convalescent sera used by the Contract Research Organization, Nexelis, during validation of the assay was used a benchmark for vaccine-induced immunogenicity. All 25 individuals had PCR-confirmed COVID-19 infection and mild to moderate symptoms. 
     In the single dose cohort, seroconversion based on antibody binding titers was observed in 12 of 16 subjects, but the titers were generally modest and significantly below those observed among convalescent sera. Neutralizing antibody responses were detected in only 8 of the 12 subjects that seroconverted and were typically just above the sensitivity of the assay. 
     Among subjects in the 2-dose cohort, there was clear evidence of boosting in both the antibody binding and neutralization titers 7 days after the second dose (Day 35), and antibody responses continued to increase from Days 28 to 56. By Day 56 all subjects had seroconverted. Immunity in this cohort was robust, with neutralization titers 4.3-fold greater than that of the convalescent sera on Day 56 (GMT of 329 vs. 76). 
     Example 12: Evaluation of Potency and Protective Effect of a Monovalent SARS-CoV-2 Beta Variant VLP Vaccine as a Booster 
     VLPs were produced using the method described Example 2 except that the SARS-CoV-2 spike protein sequence cloned into the Propol II expression vector was a SARS-CoV-2 Beta variant modified with TM/Cyt from VSV (SEQ ID NO:30). The SARS-CoV-2 sequence was from the Beta variant B 1.351 isolate EPI_ISL_911433 (GISAID). Vaccine formulations (referred to as VBI-2905a) were prepared using this Beta variant spike protein (0.1 μg) adjuvanted with aluminum phosphate adjuvant (Adjuphos®) (125 μg). Vaccine formulations referred to as VBI-2902a, described in Example 11, based on the triple modified SARs-CoV-2 Wuhan spike protein (SEQ ID: No. 24) (0.1 μg) adjuvanted with aluminum phosphate adjuvant (125 μg) were also prepared. 
     Naïve 6-8 week old C57/BL6 mice were immunized with 2 injections of VBI-2902a, 2 injections of VBI-2905a, or a first injection of VBI-2902a followed by a second injection of VBI-2905a (heterologous boost). Blood was collected on day 1 before injection and day 14 after each injection for humoral immunity assessment. 
     Anti-SARS-CoV-2 specific IgG binding titers in sera were measured by standard ELISA using recombinant SARS-CoV-2 spike RBD proteins (Sinobiological). For total IgG binding titers, detection was performed using a goat anti-mouse IgG-Fc HRP (Bethyl), or Goat anti-Hamster IgG HRP (ThermoFisher), or goat anti-human IgG heavy and light chain HRP-conjugated (Bethyl). HRP-conjugated Goat anti-mouse IgG1 and HRP-conjugated goat anti-mouse IgG2b HRP (Bethyl) were used for the detection of isotype subtype. Determination of antibody binding titers to spike RBDs was performed using a SARS-COV-2 RDB recombinant protein. The detection was completed by adding 3,3′,5,5′-tetramethylbenzidine (TMB) substrate solution, and the reaction stopped by adding liquid stop solution for TMB substrate. Absorbance was read at 450 nm in an ELISA microwell plate reader. Data fitting and analysis were performed with SoftMaxPro 5, using a four-parameter fitting algorithm. 
     Neutralizing activity in mouse serum samples was measured by standard plaque reduction neutralization test (PRNT) on Vero cells using 100 PFU of SARS-CoV2/Canada/ON/VIDO-01/2020 (Wu-1 virus) or hCoV-19/South Africa/KRISP-EC-K005321/2020 (Beta virus). Results were represented as PRNT90 end point titer (EPT), corresponding to the lowest dilution inhibiting respectively 90% of plaque formation in Vero cell culture. 
     As shown in Table 39 below, antibody binding titers to spike protein RBDs showed significant benefit of a prime vaccination with VBI-2902a and a boost with VBI-2905a. VBI-2902a induced high levels (most of the sera&gt;106 EPT with GMT 974x103) of antibody binding titers against the ancestral Wuhan spike RBD with significantly reduced cross-reactivity against the Beta variant RBD (GMT 74x103), though there was good cross-reactivity against the Delta variant RBD (GMT 616x103). Antisera from immunization with VBI-2905a showed similar cross-reactivity against the ancestral Wuhan, Delta and Beta RBD (respectively GMT 322x103, 192x103 and 217x103). However, mice receiving the heterologous prime boost regimen showed strong reactivity to each of the ancestral Wuhan and Delta RBD and to Beta RBD. 
     
       
         
           
               
             
               
                 TABLE 39 
               
             
            
               
                   
               
               
                 Antibody Binding to RBD from Different SARS-CoV-2 Strains 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 Wuhan-1 IgG 
                 Beta IgG 
                 Delta IgG 
               
               
                   
                 Mouse 
                 Wuhan-1 
                 Beta RBD 
                 Delta RBD 
                 Geometric 
                 Geometric 
                 Geometric 
               
               
                 Group 
                 # 
                 RBD IgG 
                 IgG 
                 IgG 
                 Mean 
                 Mean 
                 Mean 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 VBI-2902a 
                 25 
                 1,622,653 
                 200,759 
                 704,003 
                 974,453 
                 74,510 
                 615,553 
               
               
                   
                 26 
                 1,408,501 
                 64,036 
                 1,236,698 
               
               
                   
                 27 
                 988,821 
                 82,088 
                 734,939 
               
               
                   
                 28 
                 112,147 
                 85,278 
                 633,767 
               
               
                   
                 29 
                 2,463,443 
                 146,560 
                 810,595 
               
               
                   
                 30 
                 1,391,874 
                 59,195 
                 636,826 
               
               
                   
                 31 
                 1,049,217 
                 32,930 
                 325,804 
               
               
                   
                 32 
                 891,650 
                 36,949 
                 302,222 
               
               
                 VBI-2905a 
                 9 
                 661,450 
                 229,879 
                 318,469 
                 321,718 
                 217,804 
                 192,398 
               
               
                   
                 10 
                 555,023 
                 201,817 
                 287,502 
               
               
                   
                 11 
                 398,020 
                 294,635 
                 213,003 
               
               
                   
                 12 
                 462,292 
                 393,792 
                 303,944 
               
               
                   
                 13 
                 460,374 
                 400,407 
                 237,333 
               
               
                   
                 14 
                 56,080 
                 32,686 
                 22,659 
               
               
                   
                 15 
                 218,736 
                 169,591 
                 232,429 
               
               
                   
                 16 
                 300,841 
                 423,893 
                 253,421 
               
               
                 Heterologous 
                 49 
                 405,310 
                 71,244 
                 317,281 
                 1,045,821 
                 158,111 
                 675,623 
               
               
                 Boost 
                 50 
                 976,428 
                 260,043 
                 1,025,332 
               
               
                   
                 51 
                 233,628 
                 36,070 
                 234,033 
               
               
                   
                 52 
                 1,872,244 
                 606,640 
                 780,546 
               
               
                   
                 69 
                 2,325,516 
                 163,278 
                 1,839,676 
               
               
                   
                 70 
                 1,270,431 
                 210,707 
                 517,830 
               
               
                   
                 71 
                 2,061,624 
                 197,265 
                 884,471 
               
               
                   
                 72 
                 1,357,256 
                 141,962 
                 867,044 
               
               
                   
               
            
           
         
       
     
     Neutralizing data is shown in Table 40 below. Two doses of VBI-2902a induced high levels of neutralizing antibody response against the ancestral Wuhan strain of SARS-CoV-2 (GMT=2,458) but significantly lower activity against the Beta variant (GMT=94). By contrast, VBI-2905a induced antibodies that neutralized Beta and ancestral Wuhan viruses at similar levels in mice, yielding only a 2.2-fold difference with non significant p=0.1484. Sera from mice in the heterologous boost group cross-neutralized both the Beta variant and the ancestral Wuhan strain with similar potencies (1,4 fold difference with p=0.3828). Heterologous boosting with VBI-2905a significantly increased the PRNT90 against the ancestral strain compared to 2 doses of VBI-2905a alone (from GMT of 371 to 820, p=0.0267) to levels that were closer to those reached after two doses of VBI-2902a. 
     
       
         
           
               
             
               
                 TABLE 40 
               
             
            
               
                   
               
               
                 Neutralizing Antibodies Against Wuhan and Beta Strains of SARS-Cov-2 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Wuhan-1 
                 Beta 
                   
                   
               
               
                   
                   
                 (hCoV-19/ 
                 (hCoV-19/ 
                 Wuhan-1 
                 Beta 
               
               
                   
                   
                 Canada/ 
                 South Africa/ 
                 PRNT90 
                 PRNT90 
               
               
                   
                 Mouse 
                 ON-VIDO- 
                 KRISP-EC- 
                 Geometric 
                 Geometric 
               
               
                 Group 
                 # 
                 01/2020) 
                 K005321/2020) 
                 Mean 
                 Mean 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 VBI-2902a 
                 25 
                 5,120 
                 136 
                 2,458 
                 94 
               
               
                   
                 26 
                 3,414 
                 20 
               
               
                   
                 27 
                 853 
                 176 
               
               
                   
                 28 
                 3,149 
                 99 
               
               
                   
                 29 
                 5,120 
                 320 
               
               
                   
                 30 
                 3,149 
                 226 
               
               
                   
                 31 
                 344 
                 40 
               
               
                   
                 32 
                 5,120 
                 43 
               
               
                 VBI-2905a 
                 9 
                 640 
                 693 
                 371 
                 619 
               
               
                   
                 10 
                 320 
                 1,907 
               
               
                   
                 11 
                 800 
                 2,560 
               
               
                   
                 12 
                 452 
                 1,483 
               
               
                   
                 13 
                 427 
                 349 
               
               
                   
                 14 
                 40 
                 182 
               
               
                   
                 15 
                 686 
                 789 
               
               
                   
                 16 
                 413 
                 86 
               
               
                 Heterologous 
                 49 
                 723 
                 542 
                 820 
                 564 
               
               
                 Boost 
                 50 
                 827 
                 904 
               
               
                   
                 51 
                 320 
                 343 
               
               
                   
                 52 
                 1,721 
                 827 
               
               
                   
                 69 
                 751 
                 452 
               
               
                   
                 70 
                 640 
                 1,653 
               
               
                   
                 71 
                 827 
                 91 
               
               
                   
                 72 
                 1,566 
                 1,089 
               
               
                   
               
            
           
         
       
     
     In order to evaluate the protective effect of a heterologous boosting strategy with VBI-2905a, a challenge study was conducted in golden hamsters. Golden Syrian hamsters were intramuscularly vaccinated 3 weeks apart with two doses VBI-2902a, two doses of VBI-2905a or a priming dose of VBI-2902a followed by a second, booster dose of VBI-2905a, each formulation as described above. 
     Three weeks after the second immunization, hamsters were exposed to 1×105 TCID 50  of the Beta variant virus in each nare. In the placebo group, hamsters began losing weight the day after infection which continued until day 6-8. Vaccination with 2 doses of VBI-2902a based on the ancestral Wuhan spike protein induced limited protection against challenge with the Beta variant with moderate weight loss recorded until day 4, and only a fraction (⅗) of the animals fully regained their initial body weight after day 7. By contrast, hamsters vaccinated with 2 doses of VBI-2905a exhibited transient weight loss up to day 2-3 and then rapidly regained weight. A similar pattern was observed in hamsters that received VBI-2905a as a boost indicating that the hamsters received the same level of protection against the Beta variant from a single boost as from a two-dose regime.