Office Action 3: Notice of Allowance (2025-03-04)

CLAIMS:
LISTING OF THE CLAIMS

Although no amendments are made herein, the following listing of the claims is provided for the Examiner's convenience.

Listing of Claims:

1.-16. (Canceled)215

17\. (Previously Presented) A nucleic acid construct comprising a nucleic acid sequence encoding an exosome marker protein and a nucleic acid sequence encoding a vaccine antigen, wherein the exosome marker protein and the vaccine antigen are expressed in a fused form.

18\. (Previously Presented) The nucleic acid construct of claim 17, wherein the exosome marker protein is a protein that is present in a membrane of an exosome.

19\. (Previously Presented) The nucleic acid construct of claim 17, wherein the exosome marker protein belongs to the tetraspanin family.

20\. (Previously Presented) The nucleic acid construct of claim 17, wherein the exosome marker protein is selected from the group consisting of CD63, CD81, CD9, CD31, HLA-G, TSG101, Rab5b, and ALIX.

21\. (Previously Presented) The nucleic acid construct of claim 17, wherein the exosome marker protein is selected from the group consisting of CD63, CD81, and CD9.

22\. (Previously Presented) The nucleic acid construct of claim 17, wherein the antigen is of a pathogen.

23\. (Previously Presented) The nucleic acid construct of claim 17, wherein the nucleic acid construct is a plasmid DNA.

24\. (Previously Presented) The nucleic acid construct of claim 17, wherein the expressed protein is present in an exosome.

25\. (Previously Presented) A protein in a form of a vaccine antigen protein fused with an exosome marker protein.

26\. (Previously Presented) The protein of claim 25, wherein the exosome marker protein is a protein that is present in a membrane of an exosome.

27\. (Previously Presented) The protein of claim 25, wherein the exosome marker protein belongs to the tetraspanin family.

28\. (Previously Presented) The protein of claim 25, wherein the exosome marker protein is selected from the group consisting of CD63, CD81, CD9, CD31, HLA-G, TSG101, Rab5b, and ALIX.

29\. (Previously Presented) The protein of claim 25, wherein the exosome marker protein is selected from the group consisting of CD63, CD81, and CD9.

30\. (Previously Presented) The protein of claim 25, wherein the antigen is of a pathogen.

31\. (Previously Presented) An exosome comprising a vaccine antigen protein and an exosome marker protein in a fused form.

32\. (Previously Presented) The exosome of claim 31, wherein the exosome marker protein is a protein that is present in a membrane of an exosome.

33\. (Previously Presented) The exosome of claim 31, wherein the exosome marker protein belongs to the tetraspanin family.

34\. (Previously Presented) The exosome of claim 31, wherein the exosome marker protein is selected from the group consisting of CD63, CD81, CD9, CD31, HLA-G, TSG101, Rab5b, and ALIX.

35\. (Previously Presented) The exosome of claim 31, wherein the exosome marker protein is selected from the group consisting of CD63, CD81, and CD9.

36\. (Previously Presented) The exosome of claim 31, wherein the antigen is of a pathogen.

OFFICE ACTION:
DETAILED ACTION
Notice of Pre-AIA  or AIA  Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Application Status
This action is written in response to applicant’s correspondence received 21 January 2025. Claims 17-36 are pending. Accordingly, claims 17-36 are currently under examination.
Withdrawn Claim Rejections
Applicant’s terminal disclaimer filed 21 January 2025 has been fully considered and is
deemed persuasive. Accordingly, the double patenting rejection of record, filed 22 November
2024, has been withdrawn.
REASONS FOR ALLOWANCE
The following is an examiner’s statement of reasons for allowance: although DNA
vaccines were developed over 20 years ago, there are still concerns surrounding the efficacy of
their immunogenicity induction. Applicant has claimed a novel and non-obvious nucleic acid construct comprising a nucleic acid sequence encoding an exosome marker protein and a nucleic acid sequence encoding a vaccine antigen, wherein the exosome marker protein and the vaccine antigen are expressed in a fused form (see Claim 17).
The closest prior art, Ramos (PG Pub No. US 2017/0274062 A1), is drawn to an
invention concerned with complexes of proteins termed “Exomyc complexes” ([0001]). Ramos
teaches that the polyproteins contain immunogenic regions derived from antigens P36, P46,
HSP, NrdF, P97, MHP271 and P216 of M. hyopneumoniae, such that the regions are fused to
segments of the carrier proteins Rrp4, Rrp41 and Rrp42 of exosomes of Pyrococcus abyssi
([0011]). Ramos teaches the use of a gene or plasmid encoding the nucleotide sequences of
segments of the exosomes and the immunogenic regions such that the exosomes and
immunogenic regions are expressed in a fused form (see Claim 6).
However, Ramos does not teach or suggest that the carrier proteins are extracellular microparticles that are not degraded in the cytoplasm (Claim 17, see paragraphs [0008] and [0025]). The Exomyc complexes of Ramos is drawn to a protein complex
consisting of three copies of three different proteins that play a central role related to RNA
processing in the nuclease and cytoplasm of a cell and degrade within the cell’s cytoplasm. Accordingly, the claimed method is both novel and non-obvious in view of the closest prior art.
Conclusion

APPLICANT REBUTTAL: Application No. 18/448,713 Reply to Office Action dated November 22, 2024
REMARKS
Applicant respectfully requests reconsideration of this application in view of the above amendments and the following remarks. Claims 17-36 are pending.
Double Patenting
Claims 17-36 are provisionally rejected on the ground of nonstatutory double patenting as unpatentable over claims 1-7 of copending Application No. 17/587,925.
Claims 17-36 are provisionally rejected on the ground of nonstatutory double patenting as unpatentable over claims 11, 21-23, and 25-27 of copending Application No. 16/091,073.
To expedite prosecution without acquiescing to these rejections, Applicant files a terminal disclaimer to obviate these rejections.
Respectfully submitted, Seed Intellectual Property Law Group LLP/Qing Lin/


CITED REFERENCE SUMMARIES: Ref US20170274062A1: Claims:
A complex of immunogenic polyproteins of M. hyopneumoniae comprising immunogenic and stable segments derived from P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae, fused to segments of P. abyssi exosome, wherein the major segments of amino acids derived from M. hyopneumoniae antigens are directly linked to proteins of said P. abyssi exosome, while smaller segments are at the C-terminal extreme of the fusions.
A complex of immunogenic polyproteins according to claim 1 , wherein the proteins of P. abyssi exosome selected for fusion with the immunogenic and stable segments derived from P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae are Rrp4, Rrp41 and Rrp42, wherein the stability of the complex is reached by interaction of said proteins, thus maintaining all antigens in a single proteic complex.
A complex of immunogenic polyproteins of M. hyopneumoniae according to claim 1 , comprising an amino acids sequence selected from: (a) SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28; (b) conservatively modified variants having at least 85% of identity in relation to amino acids sequences of the sequences SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, and having substantially the same immunogenic activity thereof; (c) fragments of sequences SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, said fragments comprising immunogenic and stable regions of P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens; and (d) a fusion protein encoded by nucleotide sequence of SEQ ID NO: 25, a conservatively modified variant of said fusion protein or a fragment of said fusion protein having the same immunogenic activity.
(a) SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28;
(b) conservatively modified variants having at least 85% of identity in relation to amino acids sequences of the sequences SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, and having substantially the same immunogenic activity thereof;
(c) fragments of sequences SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, said fragments comprising immunogenic and stable regions of P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens; and
(d) a fusion protein encoded by nucleotide sequence of SEQ ID NO: 25, a conservatively modified variant of said fusion protein or a fragment of said fusion protein having the same immunogenic activity.
A complex of immunogenic polyproteins according to claim 3 , comprising the amino acids sequences of SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28.
A complex of immunogenic polyproteins according to claim 3 , comprising a fusion protein encoded by nucleotide sequence of SEQ ID NO:25.
A gene encoding a complex of immunogenic polyproteins of M. hyopneumoniae comprising nucleotide sequences encoding immunogenic and stable segments derived from P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae, and encoding nucleotide sequences of segments of P. abyssi exosome, forming a fusion protein of said immunogenic and stable segments with said segments of P. abyssi exosome so that the encoding nucleotide sequences of greater segments of amino acid derived from M. hyopneumoniae antigens are directly linked to nucleotide sequences encoding the proteins of said P. abyssi exosome, while the nucleotide sequences encoding said minor segments of amino acids derived from M. hyopneumoniae antigens are in the C-terminal extreme of the fusion protein.
A gene according to claim 6 , wherein the encoding nucleotide sequences of proteins of P. abyssi exosome selected for fusion with the immunogenic and stable segments specific of P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae are the encoding nucleotide sequences of Rrp4, Rrp41 and Rrp42 proteins of P. abyssi.
A gene encoding a complex of immunogenic polyproteins of M. hyopneumoniae according to claim 6 , comprising a nucleotide sequence selected from group consisting of: (a) SEQ ID NO:25; and (b) a nucleotide sequence having at least 75% of identity related to SEQ ID NO:25.
(a) SEQ ID NO:25; and
(b) a nucleotide sequence having at least 75% of identity related to SEQ ID NO:25.
A gene according to claim 8 , wherein said nucleotide sequence encodes the polyproteins comprising the sequences SEQ ID NO:26 SEQ ID NO:27 and SEQ ID NO:28.
A process for obtaining a complex of immunogenic polyproteins, comprising the steps of: (i) preparation of an expression system comprising: (a) a gene encoding a complex of polyproteins of M. hyopneumoniae comprising nucleotide sequences encoding immunogenic and stable segments derived from P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae and encoding nucleotide sequences of segments of P. abyssi exosome, forming a fusion protein of said immunogenic and stable segments with said segments of P. abyssi exosome so that the encoding nucleotide sequences of greater segments of amino acid derived from M. hyopneumoniae antigens are directly linked to nucleotide sequences encoding the proteins of said P. abyssi exosome, while the nucleotide sequences encoding said minor segments of amino acids derived from M. hyopneumoniae antigens are in the C-terminal extreme of the fusion protein; (b) an expression system, optionally selected from Escherichia coli strain BL21 (DE3) or Rosetta (DE3), and (c) an expression vector containing the phage promoter T7; (ii) innoculation of host cells with the expression system prepared in step (i); (iii) lysing the cells to obtain said complex of immunogenic polyproteins of M. hyopneumoniae; and (iv) purification of said complex obtained in step (iii).
(i) preparation of an expression system comprising: (a) a gene encoding a complex of polyproteins of M. hyopneumoniae comprising nucleotide sequences encoding immunogenic and stable segments derived from P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae and encoding nucleotide sequences of segments of P. abyssi exosome, forming a fusion protein of said immunogenic and stable segments with said segments of P. abyssi exosome so that the encoding nucleotide sequences of greater segments of amino acid derived from M. hyopneumoniae antigens are directly linked to nucleotide sequences encoding the proteins of said P. abyssi exosome, while the nucleotide sequences encoding said minor segments of amino acids derived from M. hyopneumoniae antigens are in the C-terminal extreme of the fusion protein; (b) an expression system, optionally selected from Escherichia coli strain BL21 (DE3) or Rosetta (DE3), and (c) an expression vector containing the phage promoter T7;
(a) a gene encoding a complex of polyproteins of M. hyopneumoniae comprising nucleotide sequences encoding immunogenic and stable segments derived from P36, P46, HSP (P42), NrdF, P97, MHP271 and P216 antigens of M. hyopneumoniae and encoding nucleotide sequences of segments of P. abyssi exosome, forming a fusion protein of said immunogenic and stable segments with said segments of P. abyssi exosome so that the encoding nucleotide sequences of greater segments of amino acid derived from M. hyopneumoniae antigens are directly linked to nucleotide sequences encoding the proteins of said P. abyssi exosome, while the nucleotide sequences encoding said minor segments of amino acids derived from M. hyopneumoniae antigens are in the C-terminal extreme of the fusion protein;
(b) an expression system, optionally selected from Escherichia coli strain BL21 (DE3) or Rosetta (DE3), and
(c) an expression vector containing the phage promoter T7;
(ii) innoculation of host cells with the expression system prepared in step (i);
(iii) lysing the cells to obtain said complex of immunogenic polyproteins of M. hyopneumoniae; and
(iv) purification of said complex obtained in step (iii).
The process according to claim 10 , wherein the encoding gene of step (i) (a) is formed by DNA amplification of primer sequences: SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO: 31 , SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36.
The process according to claim 10 , wherein said expression vector containing the phage promoter T7 of step (i) (c) is selected from the group consisting of plasmid of pET series and the pMRK plasmid.
The process according to claim 12 , wherein said plasmid of pET series is pRSET.
The process according to claim 12 , wherein said plasmid is pMRK.
An antigenic composition, comprising: (a) complex of immunogenic polyproteins of M. hyopneumoniae of claim 1 ; (b) optionally, an adjuvant; and (c) a pharmaceutically acceptable vehicle.
(a) complex of immunogenic polyproteins of M. hyopneumoniae of claim 1 ;
(b) optionally, an adjuvant; and
(c) a pharmaceutically acceptable vehicle.
The antigenic composition according to claim 15 , wherein said adjuvant is selected from group consisting of: incomplete Freund's adjuvant, the complete Freund's adjuvant, polydisperse acetylated mannan Î²-(1,4) linked, adjuvants of copolymer of polyoxiethylene-polyoxipropylene, modified lipid adjuvants, adjuvants derived from saponine, dextran sulfate, aluminum hydroxide and aluminum phosphate.
The antigenic composition according to claim 15 , wherein said pharmaceutically acceptable vehicle includes preservatives, diluents, emulsifiers, stabilizers and other pharmaceutically acceptable ingredients.
A method of treatment of infection caused by Mycoplasma hyopneumoniae, the method comprising administration of the complex of immunogenic polyproteins of M. hyopneumoniae as defined in claim 1 to a patient in need of treatment.
A method of prophylaxis of infections caused by Mycoplasma hyopneumoniae, the method comprising administration of the complex of immunogenic polyproteins of M. hyopneumoniae as defined in claim 1 to a patient in need of prophylaxis.
The process of claim 10 , wherein the gene encoding a of complex of polyproteins comprises the sequences SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28.