# Patent

## Title
Compositions and methods to immunize against hepatitis C virus

## Abstract

Compositions comprising viral antigens and antigenic peptides corresponding to or derived from Hepatitis C virus (HCV) proteins or fragments thereof, fused to heavy and/or light chain of antibodies, or fragments thereof specific for dendritic cells (DCs) are described herein. Included herein are immunostimulatory compositions (HCV vaccines, HCV antigen presenting dendritic cells, etc.) and methods for increasing effectiveness of HCV antigen presentation by an antigen presenting cell, for a treatment, a prophylaxis or a combination thereof against hepatitis C in a human subject, and methods of providing immunostimulation by activation of one or more dendritic cells, methods to treat or prevent hepatitis C.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a divisional of co-pending U.S. patent application Ser. No. 13/430,206 filed Mar. 26, 2012, which claims priority to U.S. Provisional Application Ser. No. 61/467,840, filed Mar. 25, 2011, and U.S. Provisional Application Ser. No. 61/529,700, filed Aug. 31, 2011, the entire contents of each of which are incorporated herein by reference.


TECHNICAL FIELD OF THE INVENTION
The present invention relates in general to the field of immunology, and more particularly, to hepatitis C virus (HCV) immunization, vaccines, and targeting of the HCV peptides to human dendritic cells. The application also describes a bi-functional antibody fused to a HCV target antigen(s) that is directed against a dendritic cell (DC)-specific receptor.
STATEMENT OF FEDERALLY FUNDED RESEARCH
None.
REFERENCE TO A SEQUENCE LISTING
The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Mar. 15, 2012, is named BHCS1118.txt and is 388,419 bytes in size.
BACKGROUND OF THE INVENTION
Without limiting the scope of the application, its background is described in connection with immunostimulatory methods and compositions, including vaccines, and increased effectiveness in antigen presentation of HCV peptides in relation to HCV immunization and vaccines.
U.S. Patent Application Publication No. 2009/0238822 (Rajan et al. 2009) relates to chimeric antigens for targeting and activating antigen presenting cells to elicit cellular and humoral immune responses. The Rajan invention describes compositions and methods that contain or use one or more chimeric antigens that contain one or more pre-selected HCV antigen(s), and an immunoglobulin fragment. The invention further discloses chimeric antigens, comprising an HCV antigen and a Fc fragment of an immunoglobulin for eliciting an immune response against said antigen. The immune response is said to be enhanced upon presenting the host immune system with an immune response domain (HCV antigen from HCV core, envelope, or non-structural protein fragments) and a target-binding domain (an Fc fragment).
U.S. Patent Application Publication No. 2008/0241170 (Zurawski et al. 2008) discloses compositions and methods for making and using vaccine that specifically target (deliver) antigens to antigen-presenting cells for the purpose of eliciting potent and broad immune responses directed against the antigen. The purpose is primarily to evoke protective or therapeutic immune responses against the agent (pathogen or cancer) from which the antigen was derived.
U.S. Patent Application Publication 2010/0239575 (Banchereau et al. 2010) refers to compositions and methods for the expression, secretion, and use of novel compositions for use as, e.g., vaccines and antigen delivery vectors, to delivery antigens to antigen presenting cells. In one embodiment, the vector is an anti-CD40 antibody, or fragments thereof, and one or more antigenic peptides linked to the anti-CD40 antibody or fragments thereof, including humanized antibodies.
SUMMARY OF THE INVENTION
The present invention describes immunostimulatory compositions, vaccines, HCV vaccines, HCV antigen presenting dendritic cells, methods for increasing effectiveness of HCV antigen presentation by an antigen presenting cell, methods for increasing effectiveness of HCV antigen presentation by an antigen presenting cell, methods for increasing effectiveness of antigen presentation by an antigen presenting cell, methods for a treatment, a prophylaxis or a combination thereof against hepatitis C in a human subject, methods of providing immunostimulation by activation of one or more dendritic cells, methods to treat or prevent hepatitis C in a subject, and methods for generating a HCV presenting dendritic cell. The present invention further describes virus antigens, e.g., proteins and peptides corresponding to HCV proteins or fragments thereof, fused to heavy and/or light chain of antibodies, or fragments thereof specific for dendritic cells (DCs). The vaccine composition as described herein delivers HCV antigen specifically to DCs for the purpose of invoking an immune response. The vaccine composition may also promote efficient recall memory in hepatitis C patients.
In one embodiment the instant invention discloses an immunostimulatory composition for generating an immune response for a prophylaxis, a therapy, or any combination thereof against a Hepatitis C infection in a human or animal subject comprising: one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens attached to the one or more antibodies or fragments thereof. In one aspect the composition disclosed hereinabove further comprises at least one Toll-Like Receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists. In another aspect the composition further comprises an optional pharmaceutically acceptable carrier that is effective, in combination, to produce the immune response for prophylaxis, for therapy, or any combination thereof in the human or animal subject in need of immunostimulation. In yet another aspect the DC-specific antibody or fragment is specific for a DC specific receptor, wherein the DC-specific antibody or fragment is selected from an antibody that specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, and ASPGR.
In the composition of the instant invention the HCV antigens comprises a peptide sequence derived from a HCV 1a genotype protein or a fragment thereof and the HCV antigens are selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof. The one or more HCV antigens are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and a fragment thereof and from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, E1b, and a fragment thereof. In one aspect of the composition of the instant invention the composition comprises a recombinant antibody that comprises a fusion protein and the one or more HCV antigen are at a C-terminal position relative to the one or more antibody or fragment thereof within a fusion protein. In another aspect the composition comprises a recombinant antibody, and the one or more HCV antigens are fused to a C-terminus of a heavy chain of the antibody. In yet another aspect the composition comprises a recombinant antibody, and the one or more HCV antigens are fused to a C-terminus of a light chain of the one or more antibody or fragment thereof specific for a DC.
The one or more HCV antigens are selected from the group consisting of SEQ ID NO: 12-linker A-SEQ ID NO: 13, SEQ ID NO: 12-linker A-SEQ ID NO: 11, SEQ ID NO: 12-linker B-SEQ ID NO: 14, SEQ ID NO: 14-linker B-SEQ ID NO: 12, SEQ ID NO: 12-linker B-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 12, SEQ ID NO: 9-linker
B-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 9, SEQ ID NO: 10-linker B-SEQ ID NO: 14, SEQ ID NO: 14-linker B-SEQ ID NO: 10, SEQ ID NO: 9-linker B-SEQ ID NO: 12, SEQ ID NO: 12-linker B-SEQ ID NO: 9, SEQ ID NO: 8-linker B-E1b. SEQ ID NO: 12-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 14, SEQ ID NO: 12-linker B-SEQ ID NO: 14-linker C-SEQ ID NO: 10, SEQ ID NO: 10-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 14, SEQ ID NO: 10-linker B-SEQ ID NO: 14-linker C-SEQ ID NO: 12, SEQ ID NO: 14-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 10, SEQ ID NO: 14-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 12, and SEQ ID NO: 12-linker B-SEQ ID NO: 10-linker C-SEQ ID NO: 14-linker D-SEQ ID NO: 8. In another aspect the one or more HCV antigens are attached to a C-terminus of a light chain of the recombinant antibody and selected from a group consisting of: SEQ ID NO: 9; SEQ ID NO: 11, and E1b. In yet another aspect the one or more HCV antigens are selected from the group consisting of SEQ ID NO: 9 fused to the C-terminus of a light chain and SEQ ID NO: 10-linker B-SEQ ID NO: 12-linker C-SEQ ID NO: 14 fused to the C-terminus of the heavy chain of the antibody. In a related aspect the one or more HCV antigen are chemically coupled to the one or more antibodies or fragments thereof or are attached to the one or more antibodies or fragments thereof via an affinity association. In a specific aspect the DC-specific antibody is humanized. In another aspect the composition is optimized to be administered to the human or animal subject by an oral route, a nasal route, topically, or as an injection.
Another embodiment of the present invention discloses a vaccine comprising: one or more antibodies or fragments thereof specific for a dendritic cell (DC); and one or more HCV antigens attached to the one or more antibodies or fragments thereof. The vaccine described herein further comprises at least one Toll-Like Receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists and an optional pharmaceutically acceptable carrier or an adjuvant that is effective, in combination, to produce an immune response for prophylaxis, for therapy, or any combination thereof in the human or animal subject in need of immunostimulation. In one aspect of the vaccine the DC-specific antibody or fragment is specific for a dendritic cell specific receptor. In another aspect the HCV antigen comprises a peptide sequence derived from a HCV 1a genotype protein or a fragment thereof, wherein the HCV antigen is selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof. In other related aspects the DC-specific antibody is humanized and the composition is optimized to be administered to the human or animal subject by an oral route, a nasal route, topically, or as an injection.
In yet another embodiment the instant invention discloses a Hepatitis C vaccine (HCV) comprising a fusion protein comprising: (i) one or more antibodies or fragments thereof specific for a dendritic cell (DC), (ii) one or more HCV antigens located C-terminal of the antibodies or fragments thereof, (iii) at least one Toll-Like Receptor (TLR) agonist which is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists, and (iv) one or more optional pharmaceutically acceptable carriers and adjuvants, wherein the vaccine is effective to produce an immune response, for a prophylaxis, a therapy, or any combination thereof against hepatitis C in a human or an animal subject in need thereof. In one aspect the vaccine comprises one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof.
The instant invention in one embodiment discloses a method for increasing effectiveness of Hepatitis C virus (HCV) antigen presentation by an antigen presenting cell (APC) comprising the steps of: (i) providing an antibody conjugate comprising a dendritic cell (DC) specific antibody or a fragment thereof and one or more native or engineered HCV antigenic peptides, (ii) providing one or more APCs; and (iii) contacting the APC with the conjugate, wherein the antibody-antigen complex is processed and presented for T cell recognition. In a specific aspect of the method the antigen presenting cell comprises a dendritic cell (DC).
In another embodiment the instant invention provides a method for increasing effectiveness of antigen presentation by an antigen presenting cell (APC) comprising the steps of: i) isolating and purifying one or more dendritic cell (DC)-specific antibody or a fragment thereof, ii) providing one or more HCV antigens or antigenic peptides, iii) loading or chemically coupling the one or more HCV antigens or antigenic peptides to the DC-specific antibody to form an antibody-antigen conjugate, and iv) contacting the antigen presenting cell with the conjugate, wherein the antibody-antigen complex is processed and presented for T cell recognition.
The method as described hereinabove further comprises adding at least one Toll-Like Receptor (TLR) agonist which is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists and one or more optional steps comprising i) adding one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof to the antibody-antigen conjugate and the TLR agonist prior to contacting the antigen presenting cells, ii) measuring a level of one or more agents selected from the group consisting of IFN-γ, TNF-α, IL-12p40, IL-4, IL-5, and IL-13, wherein a change in the level of the one or more agents is indicative of the increase in the effectiveness antigen presentation by the antigen presenting cell, and iii) adding one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof.
In yet another embodiment the instant invention provides method for a treatment, a prophylaxis or a combination thereof against hepatitis C in a human subject comprising the steps of: identifying the human subject in need of the treatment, the prophylaxis, or a combination thereof against the hepatisti and administering a vaccine composition comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens attached to the one or more antibodies or fragments thereof. In one aspect of the method the vaccine composition further comprises at least one Toll-Like Receptor (TLR) agonist which is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists, and one or more optional pharmaceutically acceptable carriers and adjuvants, wherein the conjugate and agonist are each comprised in an amount such that, in combination with the other, are effective to produce an immune response, for the prophylaxis, the therapy or any combination thereof against the influenza in the human subject. In another aspect the vaccine composition further comprises one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof. In yet another aspect the vaccine is administered to the human subject by an oral route, a nasal route, topically or as an injection.
In another aspect the one or more antibodies or fragments thereof specific for a dendritic cell comprises antibodies specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fey receptor, LOX-1, or ASPGR. In yet another aspect the HCV antigen is selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof, from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and a fragment thereof, or from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, E1b, and a fragment thereof.
A method of providing immunostimulation by activation of one or more dendritic cells (DCs) to a human subject for a prophylaxis, a therapy or a combination thereof against HCV is described in one embodiment of the present invention. The method comprises the steps of: a) identifying the human subject in need of immunostimulation for the prophylaxis, the therapy or a combination thereof against HCV, b) isolating one or more DCs from the human subject, c) exposing the isolated DCs to activating amounts of a composition or a vaccine comprising an anti-dendritic cell immunoreceptor (DCIR) monoclonal antibody or fragments thereof attached to one or more HCV antigens, and d) reintroducing the activated DC complex into the human subject.
The method described above further comprises the steps of contacting the one or more DCs with at least one Toll-Like Receptor (TLR) agonist which is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists and a pharmaceutically acceptable carrier to form an activated DC complex and the step of adding one or more optional agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines or combinations and modifications thereof to the conjugate and the TLR agonist prior to exposing the DCs. The method disclosed hereinabove further comprises the optional step of measuring a level of one or more agents selected from the group consisting of IFN-γ, TNF-α, IL-12p40, IL-4, IL-5, and IL-13, wherein a change in the level of the one or more agents is indicative of the immunostimulation.
The present invention also discloses a method to treat or prevent Hepatitis C in a subject comprising the step of administering to the subject a fusion protein comprising an antibody or fragment thereof specific for a dendritic cell (DC) and a Hepatitis C virus antigen or antigenic peptide fused to the antibody or fragment thereof. A Hepatitis C virus antigen presenting dendritic cell (DC) is also disclosed in one embodiment of the present invention. The HCV antigen presenting DC further comprises one or more isolated dendritic cells (DCs) in contact with a fusion protein comprising an antibody or fragment thereof specific for the DC, the fusion protein further comprising a HCV peptide.
The present invention describes one or more vaccines against HCV comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens or antigenic domains attached to the one or more antibodies or fragments thereof. The vaccine has a general structure given by: H-w, H-w-x, H-w-x-y, or H-w-x-y-z, wherein H represents a heavy chain of an antibody or a fragment thereof specific for a DC, w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof. In one aspect w comprises the HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, Palm, E1b, and E2. In another aspect x comprises the HCV antigenic domains selected from the group consisting of Hel C, Hel A, Palm, ProtA, ProtB, and E1b. In yet another aspect comprises the HCV antigenic domains selected from the group consisting of Palm, ProtB, and Protb. In another aspect z comprises HCV antigenic domains selected from E2, ProtA, and HelB. In a related aspect the one or more HCV antigens or antigenic domains are linked or attached to one another by one or more flexible linkers.
Another embodiment disclosed herein relates to a vaccine comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens or antigenic domains attached to the one or more antibodies or fragments thereof, wherein the vaccine has a general structure given by L-w-x-y-z, wherein L represents a light chain of an antibody or a fragment thereof specific for a DC, w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof.
In yet another embodiment the present invention discloses a vaccine comprising one or more antibodies or fragments thereof specific for a dendritic cell (DC) and one or more HCV antigens or antigenic domains attached to the one or more antibodies or fragments thereof, wherein the vaccine has a general structure given by:




Wherein H represents a heavy chain of an antibody or a fragment thereof specific for a DC, L represents a light chain of an antibody or a fragment thereof specific for the DC, w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof.
Finally, the present invention discloses a method for generating a Hepatitis C virus (HCV) presenting dendritic cells (DCs) in a human subject comprising the steps of: providing one or more DCs and incubating the dendritic cells with a fusion protein, wherein the fusion protein comprises an antibody or fragment thereof specific for a dendritic cell and a HCV antigen fused to the antibody or fragment thereof. The method disclosed herein further comprises the step of administering to the subject an effective amount of IFNA, Ribavirin, or a combination thereof.



BRIEF DESCRIPTION OF THE DRAWINGS
For a more complete understanding of the features and advantages of the present invention, reference is now made to the detailed description of the invention along with the accompanying figures and in which:
FIG. 1 provides a summary of HCV antigen combined constructs expressed at the C-terminus end of antibody heavy chain. Each HCV domains, as defined in FIG. 2, is represented as color rectangle. Flexible linkers are shown as curved lines. Each color represents a different linker. Domains are fused to the carboxyl terminus end of antibody heavy chain. “Expressed” means that domains fused to the carboxyl terminus end of antibody heavy chain are expressed as fusion antibody after co-transfection with antibody light chain in 293F cells. All possible combination of HCV domains have been constructed, and FIG. 3 shows only those that were expressed as soluble fusion proteins in 293F cells and in CHO cells and purified as recombinant antibodies;
FIG. 2 provides a summary of HCV antigen combined constructs expressed at the C-terminus end of antibody light chain. All possible combinations of HCV domains have been constructed, and the figure shows only those that are expressed as soluble fusion proteins in 293F cells and in CHO cells and purified as recombinant antibodies. The same color code as in FIG. 1 is used;
FIGS. 3A-3B demonstrate the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB specific antigen to elicit the expansion of antigen-specific CD4+ T cells from a chronic HCV infected patient cured after IFNa-Ribavirin therapy. Delivering NS3HelB to DCs through CD40 and DCIR induces IFNγ-TNFa-producing HCV NS3HelB-specific CD4+ T cells. PBMC cells from chronic HCV infected patients; either cured after therapy or in treatment failure, were co-cultured with IFNDCs targeted with anti-CD40-NS3HelB or anti-DCIR-NS3HelB for 10 days. Cells were stimulated with peptides clusters (10 peptides of 15-mers in each clusters) covering HCV NS3 HelB (10 μM): FIG. 3A after 2 days, culture supernatants were analyzed for measuring IFNγ and FIG. 3B PBMC cells were stained for measuring the frequency of peptide-specific CD4+ T cells intracellular IFNγ+TNFα+ cells;
FIG. 4 demonstrates the ability of long HCV antigen bearing vaccine constructs to induce multi epitope CD4+ T cells. HCV antigens from NS3 Helicase HelBC construct were delivered to DCs through CD40 or DCIR. PBMC cells from chronic HCV infected patients; either cured after therapy or in treatment failure, were co-cultured with IFNDCs targeted with anti-CD40-NS3HelB, anti-CD40-NS3HelBC or anti-DCIR-NS3HelB, anti-DCIR-NS3HelBC for 10 days. Cells were stimulated for 6 h with peptides clusters (10 μM; 10 peptides of 15-mers in each clusters) covering HCV NS3 HelB or HelBC constructs. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ T cells intracellular IFNγ+TNFα+ cells, an analyzed by FACS. Number of double positive CD4+ T cells induced after each peptide cluster stimulation were plotted for each vaccine targeting agent;
FIGS. 5A to 5C demonstrate the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB, HCV NS3ProtB and HCV NS5BPalm specific antigens to elicit the expansion of antigen-specific CD4+ T cells from a chronic HCV infected patient cured after IFNa-Ribavirin therapy. Delivering HCV antigen to DCs through CD40 and DCIR induces IFNγ-TNFa-producing HCV-specific CD4+ T cells, with multi epitopes, specific CD4 T cells. PBMC cells from chronic HCV infected patients cured after therapy were co-cultured with autologous IFNαDCs targeted with anti-CD40-NS3HelB-NS3ProtB-NS5BPalm or anti-DCIR-NS3HelB-NS3ProtB-NS5BPalm for 10 days. Cells were stimulated with peptides clusters (10 peptides of 15-mers in each clusters) covering HCV NS3HelB, NS3ProtB or NS5BPalm (2 μM). PBMC cells were stained for measuring the frequency of peptide-specific CD4+ T cells intracellular IFNγ+TNFα+ cells;
FIGS. 6A to 6C demonstrate the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB, HCV NS3ProtB and HCV NS5BPalm specific antigens to elicit the expansion of antigen-specific CD8+ T cells from a chronic HCV infected patient cured after IFNa-Ribavirin therapy. Delivering HCV antigen to DCs through CD40 and DCIR induces IFNγ-TNFa-producing HCV-specific CD4+ T cells, with multi epitopes, specific CD4 T cells. PBMC cells from chronic HCV infected patients cured after therapy were co-cultured with autologous IFNαDCs targeted with anti-CD40-NS3HelB-NS3ProtB-NS5BPalm or anti-DCIR-NS3HelB-NS3ProtB-NS5BPalm for 10 days. Cells were stimulated with peptides clusters (10 peptides of 15-mers in each clusters) covering HCV NS3HelB, NS3ProtB or NS5BPalm (2 μM). PBMC cells were stained for measuring the frequency of peptide-specific CD8+T cells intracellular IFNγ+TNFα+ cells;
FIGS. 7A to 7D demonstrate the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB, HCV NS3ProtB or HCV NS5BPalm specific antigens to elicit the expansion of antigen-specific CD4+ T cells from chronic HCV infected patients cured after IFNa-Ribavirin therapy. HCV antigens from were delivered to DCs through CD40 or DCIR. IFNαDCs were targeted with anti-CD40-NS3HelB; anti-CD40-NS3ProtB; anti-CD40NS5bPalm or anti-DCIR-NS3HelB; anti-DCIR-NS3ProtB; anti-DCIRNS5bPalm and co-cultured for 10 days with PBMC cells from 3 chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7 and C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB constructs; with peptide clusters C2 and C3 covering HCV NS3 ProtB constructs or peptide cluster C2 C4 C5 C6 C7 covering NS5bPalm construct. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS. The left panel represent IFNγ amount secreted after 10 days culture of PBMCs with peptide cluster covering HCVNS3 and HCVNS5b entire proteins;
FIGS. 8A to 8D demonstrates the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB, HCV NS3ProtB or HCV NS5BPalm specific antigens to elicit the expansion of antigen-specific CD4+ T cells from chronic HCV infected patients in treatment failure. HCV antigens from were delivered to DCs through CD40 or DCIR. IFNαDCs were targeted with anti-CD40-NS3HelB; anti-CD40-NS3ProtB or anti-DCIR-NS3HelB; anti-CD40NS5bPalm or anti-DCIR-NS3HelB; anti-DCIR-NS3ProtB; anti-DCIRNS5bPalm and co-cultured for 10 days with PBMC cells from 3 chronic HCV infected patients in treatment failure (HCV+). Cells were stimulated for 6 h with peptide clusters C7 and C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB constructs; with peptide clusters C2 and C3 covering HCV NS3 ProtB constructs or peptide cluster C2 C4 C5 C6 C7 covering NS5bPalm construct. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS. The left panel represent IFNγ amount secreted after 10 days culture of PBMCs with peptide cluster covering HCVNS3 and HCVNS5b entire proteins;
FIGS. 9A and 9B demonstrate the ability of combination of TLR agonist and anti-DCIR HCV-NS3HelB can induced multi epitopes CD8+ T cells. HCV antigens from NS3 Helicase HelB construct were delivered to DCs through CD40 or DCIR. IFNDCs were targeted with anti-CD40-NS3HelB, or anti-DCIR-NS3HelB in presence of PAM3 (TLR2 agonist; 200 ng/ml), CL097 (TLR7/8 agonist; 5 μg/ml) or polyIC (TLR3 agonist; 25 μg/ml) before co-culture for 10 days with PBMC cells from chronic HCV infected patients; either cured after therapy or in treatment failure. Cells were stimulated for 6 h with peptide clusters C7 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB constructs: (FIG. 9A) PBMC cells were stained for measuring the frequency of peptide-specific CD4+ and CD8+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS, (FIG. 9B) Number of double positive CD4+ and CD8+ intracellular IFNγ+TNFα+ T cells induced after each TLR agonist stimulation were plotted;
FIGS. 10A-10D demonstrate the ability of combination of TLR agonist and anti-DCIR HCV-construct to increase CD4+ and induce CD8+ T cells responses in chronic HCV infected patients cured after therapy. HCV antigens from NS3 Helicase HelB or from NS3 Protease ProtB constructs were delivered to DCs through CD40 or DCIR. IFNαDCs were targeted with anti-CD40-NS3HelB, anti-DCIR-NS3HelB, anti-CD40-NS3ProtB, anti-DCIR-NS3ProtB, in presence of PAM3 (TLR2 agonist; 200 ng/ml), CL095 (TLR7/8 agonist; 5 μg/ml) or polyIC (TLR3 agonist; 25 μg/ml) or cyclic glucan (TLR4 agonist, 10 m/ml) before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB constructs or with peptide clusters C3 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB constructs: FIGS. 10A and 10B PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, and analyzed by FACS and FIGS. 10C and 10D PBMC cells were stained for measuring the frequency of peptide-specific CD8+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS;
FIG. 11 demonstrates the ability of combination of TLR agonists and anti-CD40 HCV-constructs to increase CD4+ T cells responses in chronic HCV infected patients in treatment failure. HCV antigens from NS3 Helicase HelB or from NS3 Protease ProtB constructs were delivered to DCs through CD40 or DCIR. IFNαDCs were targeted with anti-CD40-NS3HelB, anti-DCIR-NS3HelB, anti-CD40-NS3ProtB, anti-DCIR-NS3ProtB, in presence of PAM3 (TLR2 agonist; 200 ng/ml), CL095 (TLR7/8 agonist; 5 μg/ml) or polyIC (TLR3 agonist; 25 μg/ml) or cyclic glucan (TLR4 agonist, 10 μg/ml) before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB constructs or with peptide clusters C3 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB constructs. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS;
FIGS. 12A-12C demonstrate the ability of HCV vaccine candidates to recall CD4+ T cells responses in all chronic HCV infected patients (cured or in treatment failure). HCV antigens from NS3 Helicase HelB, NS5b polymerase Palm or from NS3 Protease ProtB constructs were delivered to DCs through CD40 or DCIR. IFNγDCs were targeted with anti-CD40-[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain], anti-DCIR-[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain] before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7, C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB domain, with peptide clusters C2-C3-C4 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB domain or with peptide clusters C2-C4-C5-C6-C7 (10 μM; 10 peptides of 15-mers) covering HCV NS5b Palm domain. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ and CD8+ intracellular IFNγ+TNFα+ cells, and analyzed by FACS. The number of CD4+IFNγ+TNFα+ cells induced vaccine candidate is shown;
FIGS. 13A-13E demonstrate of the ability of different HCV antigen combination on vaccine candidate for recall CD4+ T cells responses in chronic HCV infected cured patients. HCV antigens from NS3 Helicase HelB, NS5b polymerase Palm or from NS3 Protease ProtB combination constructs were delivered to DCs through CD40 or DCIR. IFNaDCs were targeted with second-generation vaccines anti-CD40-[NS3HelB on light chain and NS3ProtB ̃NS5bPalm on heavy chain], anti-DCIR-[NS3HelB on light chain and NS3ProtB ̃ NS5bPalm on heavy chain], or first-generation vaccines anti-CD40-[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain], anti-DCIR—[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain] before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7 and C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB domain (shown in green on the figure), with peptide clusters C2-C3-C4 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB domain (shown in pink on the figure) or with peptide clusters C2-C4-C5-C6-C7 (10 μM; 10 peptides of 15-mers) covering HCV NS5b Palm domain (shown in orange in the figure). PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFa+ cells, an analyzed by FACS. The number of CD4+IFNγ+TNFα+ cells induced by first-generation vaccine or second-generation vaccine is compared in the last panel; and
FIGS. 14A to 14H demonstrate the ability of vaccine candidate to recall CD4+ T cells responses in HCV patients infected with non 1 genotype and HCV-exposed but non-infected individual. HCV antigens from NS3 Helicase HelB, NS5b polymerase Palm or from NS3 Protease ProtB combination constructs were delivered to DCs through CD40 or DCIR and DC loaded were co-culture for 10 days with PBMC cells from HCV patients infected with non 1 genotype HCV-infected patients (HCV-015, 2b) and HCV-exposed but non infected individual (HCV-029). Cells were stimulated for 6 h with peptide clusters C7 and C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB domain, with peptide clusters C2-C3-C4 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB domain or with peptide clusters C2-C4-C5-C6-C7 (10 μM; 10 peptides of 15-mers) covering HCV NS5b Palm domain. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS.
FIGS. 15A to 15B shows the results from a 10 day expansion culture whereby a dose range of 1st generation anti-DCIR-HCV vaccine (left panels) is compared to second generation anti-DCIR-HCV vaccine (right panels). Doses were 0.05 nM, 0.5 nM, and 5 nM and antigen-specific responses were ascertained by stimulation with no peptide (control) or ProtA, HelB, or Palm peptide pools in the presence of Brefeldin, followed by staining for CD3+, CD4+ and intracellular IFNg and TNFa. Samples were analyzed by FACS. Shown are comparable CD4+ HCV antigen-specific responses to the two generations of vaccines.
FIGS. 16A to 16B shows the results from a 10 day expansion culture whereby a dose range of 1st generation anti-CD40-HCV vaccine (left panels) is compared to second generation anti-CD40-HCV vaccine (right panels). Doses were 0.05 nM, 0.5 nM, and 5 nM and antigen-specific responses were ascertained by stimulation with no peptide (control) or ProtA, HelB, or Palm peptide pools in the presence of Brefeldin, followed by staining for CD3+, CD4+ and intracellular IFNg and TNFa. Samples were analyzed by FACS. Shown are comparable CD4+ HCV antigen-specific responses to the two generations of vaccines.



DETAILED DESCRIPTION OF THE INVENTION
While the making and using of various embodiments of the present invention are discussed in detail below, it should be appreciated that the present invention provides many applicable inventive concepts that can be embodied in a wide variety of specific contexts. The specific embodiments discussed herein are merely illustrative of specific ways to make and use the invention and do not delimit the scope of the invention.
To facilitate the understanding of this invention, a number of terms are defined below. Terms defined herein have meanings as commonly understood by a person of ordinary skill in the areas relevant to the present invention. Terms such as “a”, “an” and “the” are not intended to refer to only a singular entity, but include the general class of which a specific example may be used for illustration. The terminology herein is used to describe specific embodiments of the invention, but their usage does not delimit the invention, except as outlined in the claims.
The invention includes also variants and other modification of an antibody (or “Ab”) of fragments thereof, e.g., anti-CD40 fusion protein (antibody is used interchangeably with the term “immunoglobulin”). As used herein, the term “antibodies or fragments thereof,” includes whole antibodies or fragments of an antibody, e.g., Fv, Fab, Fab′, F(ab′)2, Fc, and single chain Fv fragments (ScFv) or any biologically effective fragments of an immunoglobulins that binds specifically to, e.g., CD40. Antibodies from human origin or humanized antibodies have lowered or no immunogenicity in humans and have a lower number or no immunogenic epitopes compared to non-human antibodies. Antibodies and their fragments will generally be selected to have a reduced level or no antigenicity in humans.
As used herein, the terms “Ag” or “antigen” refer to a substance capable of either binding to an antigen binding region of an immunoglobulin molecule or of eliciting an immune response, e.g., a T cell-mediated immune response by the presentation of the antigen on Major Histocompatibility Antigen (MHC) cellular proteins. As used herein, “antigen” includes, but is not limited to, antigenic determinants, haptens, and immunogens, which may be peptides, small molecules, carbohydrates, lipids, nucleic acids or combinations thereof. The skilled immunologist will recognize that when discussing antigens that are processed for presentation to T cells, the term “antigen” refers to those portions of the antigen (e.g., a peptide fragment) that is a T cell epitope presented by MHC to the T cell receptor. When used in the context of a B cell mediated immune response in the form of an antibody that is specific for an “antigen”, the portion of the antigen that binds to the complementarity determining regions of the variable domains of the antibody (light and heavy) the bound portion may be a linear or three-dimensional epitope. In the context of the present invention, the term antigen is used on both contexts, that is, the antibody is specific for a protein antigen (CD40), but also carries one or more peptide epitopes for presentation by MHC to T cells. In certain cases, the antigens delivered by the vaccine or fusion protein of the present invention are internalized and processed by antigen presenting cells prior to presentation, e.g., by cleavage of one or more portions of the antibody or fusion protein.
As used herein, the term “conjugate” refers to a protein having one or more targeting domains, e.g., an antibody, and at least one antigen, e.g., a small peptide or a protein. These conjugates include those produced by chemical methods, such as by chemical coupling, for example, coupling to sulfhydryl groups, and those produced by any other method whereby one or more antibody targeting domains and at least one antigen, are linked, directly or indirectly via linker(s) to a targeting agent. An example of a linker is a cohesin-dockerin (coh-doc) pair, a biotin-avidin pair, histidine tags bound by Zn, and the like.
As used herein, the term “Antigen Presenting Cells” (APC) refers to cells that are capable of activating T cells, and include, but are not limited to, certain macrophages, B cells and dendritic cells. “Dendritic cells” (DCs) refers to any member of a diverse population of morphologically similar cell types found in lymphoid or non-lymphoid tissues. These cells are characterized by their distinctive morphology, high levels of surface MHC-class II expression (Steinman, et al., Ann. Rev. Immunol. 9:271 (1991); incorporated herein by reference for its description of such cells). These cells can be isolated from a number of tissue sources, and conveniently, from peripheral blood, as described herein. Dendritic cell binding proteins refers to any protein for which receptors are expressed on a dendritic cell. Examples include GMCSF, IL-1, TNF, IL-4, CD40L, CTLA4, CD28, and FLT-3 ligand.
For the purpose of the present invention, the term “vaccine composition” is intended to mean a composition that can be administered to humans or to animals in order to induce an immune system response; this immune system response can result in a production of antibodies or simply in the activation of certain cells, in particular antigen-presenting cells, T lymphocytes and B lymphocytes. The vaccine composition can be a composition for prophylactic purposes or for therapeutic purposes, or both. As used herein, the term “antigen” refers to any antigen which can be used in a vaccine, whether it involves a whole microorganism or a subunit, and whatever its nature: peptide, protein, glycoprotein, polysaccharide, glycolipid, lipopeptide, etc. They may be viral antigens, bacterial antigens, or the like; the term “antigen” also comprises the polynucleotides, the sequences of which are chosen so as to encode the antigens whose expression by the individuals to which the polynucleotides are administered is desired, in the case of the immunization technique referred to as DNA immunization. They may also be a set of antigens, in particular in the case of a multivalent vaccine composition which comprises antigens capable of protecting against several diseases, and which is then generally referred to as a vaccine combination, or in the case of a composition which comprises several different antigens in order to protect against a single disease, as is the case for certain vaccines against whooping cough or the flu, for example. The term “antibodies” refers to immunoglobulins, whether natural or partially or wholly produced artificially, e.g. recombinant. An antibody may be monoclonal or polyclonal. The antibody may, in some cases, be a member of one, or a combination immunoglobulin classes, including: IgG, IgM, IgA, IgD, and IgE.
The term “adjuvant” refers to a substance that enhances, augments or potentiates the host's immune response to a vaccine antigen.
The term “gene” is used to refer to a functional protein, polypeptide or peptide-encoding unit. As will be understood by those in the art, this functional term includes both genomic sequences, cDNA sequences, and fragments or combinations thereof, as well as gene products, including those that may have been altered by the hand of man. Purified genes, nucleic acids, protein and the like are used to refer to these entities when identified and separated from at least one contaminating nucleic acid or protein with which it is ordinarily associated.
As used herein, the term “nucleic acid” or “nucleic acid molecule” refers to polynucleotides, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments generated by the polymerase chain reaction (PCR), and fragments generated by any of ligation, scission, endonuclease action, and exonuclease action. Nucleic acid molecules can be composed of monomers that are naturally-occurring nucleotides (such as DNA and RNA), or analogs of naturally-occurring nucleotides (e.g., α-enantiomeric forms of naturally-occurring nucleotides), or a combination of both. Modified nucleotides can have alterations in sugar moieties and/or in pyrimidine or purine base moieties. Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyl groups, amines, and azido groups, or sugars can be functionalized as ethers or esters. Moreover, the entire sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs. Examples of modifications in a base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substitutes. Nucleic acid monomers can be linked by phosphodiester bonds or analogs of such linkages. Analogs of phosphodiester linkages include phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoranilidate, phosphoramidate, and the like. The term “nucleic acid molecule” also includes so-called “peptide nucleic acids,” which comprise naturally-occurring or modified nucleic acid bases attached to a polyamide backbone. Nucleic acids can be either single stranded or double stranded.
As used herein, “polynucleotide” or “nucleic acid” refers to a strand of deoxyribonucleotides or ribonucleotides in either a single- or a double-stranded form (including known analogs of natural nucleotides). A double-stranded nucleic acid sequence will include the complementary sequence. The polynucleotide sequence may encode variable and/or constant region domains of immunoglobulin that are formed into a fusion protein with one or more linkers. For use with the present invention, multiple cloning sites (MCS) may be engineered into the locations at the carboxy-terminal end of the heavy and/or light chains of the antibodies to allow for in-frame insertion of peptide for expression between the linkers. As used herein, the term “isolated polynucleotide” refers to a polynucleotide of genomic, cDNA, or synthetic origin or some combination thereof. By virtue of its origin the “isolated polynucleotide” (1) is not associated with all or a portion of a polynucleotide in which the “isolated polynucleotides” are found in nature, (2) is operably linked to a polynucleotide which it is not linked to in nature, or (3) does not occur in nature as part of a larger sequence. The skilled artisan will recognize that to design and implement a vector can be manipulated at the nucleic acid level by using techniques known in the art, such as those taught in Current Protocols in Molecular Biology, 2007 by John Wiley and Sons, relevant portions incorporated herein by reference. Briefly, the encoding nucleic acid sequences can be inserted using polymerase chain reaction, enzymatic insertion of oligonucleotides or polymerase chain reaction fragments in a vector, which may be an expression vector. To facilitate the insertion of inserts at the carboxy terminus of the antibody light chain, the heavy chain, or both, a multiple cloning site (MCS) may be engineered in sequence with the antibody sequences.
As used herein, the term “polypeptide” refers to a polymer of amino acids and does not refer to a specific length of the product; thus, peptides, oligopeptides, and proteins are included within the definition of polypeptide. This term also does not refer to or exclude post expression modifications of the polypeptide, for example, glycosylations, acetylations, phosphorylations and the like. Included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), polypeptides with substituted linkages, as well as other modifications known in the art, both naturally occurring and non-naturally occurring. The term “domain,” or “polypeptide domain” refers to that sequence of a polypeptide that folds into a single globular region in its native conformation, and that may exhibit discrete binding or functional properties.
As used in this application, the term “amino acid” means one of the naturally occurring amino carboxylic acids of which proteins are comprised. The term “polypeptide” as described herein refers to a polymer of amino acid residues joined by peptide bonds, whether produced naturally or synthetically. Polypeptides of less than about 10 amino acid residues are commonly referred to as “peptides.” A “protein” is a macromolecule comprising one or more polypeptide chains. A protein may also comprise non-peptidic components, such as carbohydrate groups. Carbohydrates and other non-peptidic substituents may be added to a protein by the cell in which the protein is produced, and will vary with the type of cell. Proteins are defined herein in terms of their amino acid backbone structures; substituents such as carbohydrate groups are generally not specified, but may be present nonetheless.
A polypeptide or amino acid sequence “derived from” a designated nucleic acid sequence refers to a polypeptide having an amino acid sequence identical to that of a polypeptide encoded in the sequence, or a portion thereof wherein the portion consists of at least 3-5 amino acids, preferably at least 4-7 amino acids, more preferably at least 8-10 amino acids, and even more preferably at least 11-15 amino acids, or which is immunologically identifiable with a polypeptide encoded in the sequence. This terminology also includes a polypeptide expressed from a designated nucleic acid sequence.
As used herein, the terms “stable,” “soluble,” or “unstable” when referring to proteins is used to describe a peptide or protein that maintains its three-dimensional structure and/or activity (stable) or that loses immediately or over time its three-dimensional structure and/or activity (unstable). As used herein, the term “insoluble” refers to those proteins that when produced in a cell (e.g., a recombinant protein expressed in a eukaryotic or prokaryotic cell or in vitro) are not soluble in solution absent the use of denaturing conditions or agents (e.g., heat or chemical denaturants, respectively). The antibody or fragment thereof and the linkers taught herein have been found to convert antibody fusion proteins with the peptides from insoluble and/or unstable into proteins that are stable and/or soluble. Another example of stability versus instability is when the domain of the protein with a stable conformation has a higher melting temperature (Tm) than the unstable domain of the protein when measured in the same solution. A domain is stable compared to another domain when the difference in the Tm is at least about 2° C., more preferably about 4° C., still more preferably about 7° C., yet more preferably about 10° C., even more preferably about 15° C., still more preferably about 20° C., even still more preferably about 25° C., and most preferably about 30° C., when measured in the same solution.
As used herein, the term “in vivo” refers to being inside the body. The term “in vitro” used as used in the present application is to be understood as indicating an operation carried out in a non-living system.
As used herein, the term “treatment” or “treating” means any administration of a compound of the present invention and includes (1) inhibiting the disease in an animal that is experiencing or displaying the pathology or symptomatology of the diseased (i.e., arresting further development of the pathology and/or symptomatology), or (2) ameliorating the disease in an animal that is experiencing or displaying the pathology or symptomatology of the diseased (i.e., reversing the pathology and/or symptomatology).
As used herein, “pharmaceutically acceptable carrier” refers to any material that when combined with an immunoglobulin (Ig) fusion protein of the present invention allows the Ig to retain biological activity and is generally non-reactive with the subject's immune system. Examples include, but are not limited to, standard pharmaceutical carriers such as a phosphate buffered saline solution, water, emulsions such as an oil/water emulsion, and various types of wetting agents. Certain diluents may be used with the present invention, e.g., for aerosol or parenteral administration, that may be phosphate buffered saline or normal (0.85%) saline.
Substantial similarity of a peptide refers to similarity of a peptide as reflected in the amino acid sequence of the peptide. Identity of a continuous stretch of least 8 amino acids in an antigenic epitope of the peptide may be sufficient to establish substantial identity that enables cross reactivity. A first peptide and a second peptide are substantially similar in this regard if they have substantial similar antigenic epitopes so that immunization with the first peptide causes an immune response against the second peptide.
A fragment of an antibody, as used in the present application, refers to a portion of an antibody, created by protein engineering including proteolysis, or genetic engineering including recombination of nucleic acids; the fragment of an antibody retains specificity for the antigen.
A fragment of a peptide used as antigen refers to a portion of the peptide that retains its immunogenicity. A person of ordinary skill in the art will recognize that a continuous stretch of least 8 amino acids in an antigenic epitope of the peptide may be sufficient I order for a peptide to retain its immunogenicity.
Recombinant protein or antibody is generated by genetic engineering of nucleic acid encoding the protein or antibody and subsequent translation of the coding sequence by a cell or in a cell-free translation system.
The present invention describes a vaccine composition for delivering a HCV antigen specifically to DCs for the purpose of invoking an immune response In one embodiment, due to the high polymorphism of HCV, a sequence that is representative of most of circulating HCV sequence was selected. Based on sequence variation HCV can be classified into 6 genotypes that differs one to the other on the basis of sequence identity. World wide, 1 genotype is the most represented and also the most difficult to treat with the current IFNa-Ribavirin double therapy. More precisely, 1a genotype is the most represented subsequence in industrial country, and especially in US.
In one embodiment, 1a genotype was used as target sequence to derive a vaccine. It was observed that sequence alignment with all available 1a sequences found in data bases (euHCVdb and Los Alamos National Laboratory) showed less than 70% of sequence identity and the sequence of the HCV antigen would have to be adjusted accordingly.
A mosaic sequence was derived using the mosaic vaccine tools at www.hiv.lanl.gov/content/sequence/MOSAIC/ interface choosing mosaic sequence cocktail, 1 as cocktail size and 9 as epitope size. We used 249 sequences for E1 mosaic, 656 sequences for E2 mosaic, 213 sequences for NS3 mosaic and 310 sequences for NS5b mosaic. All sequences correspond to complete genes of 576, 1089, 1893, 1773 nucleotides respectively and found in euHCVdb (euhcvdb.ibcp.fr/euHCVdb/).
HCV antigen choice: HCV is an RNA enveloped virus. Virions are consisted by 4 structural proteins Core, E1, E2 and p7. As an RNA virus replication is based on viral proteins that need to be expressed after infection. Six non-structural proteins (NS2, NS3, NS4a, NS4b, NS5a, NS5b) are necessary to establish and maintain replication and virus production. HCV targets the liver and can infect barely all the liver with 90% of hepatocytes infected. However, the virus is able to replicate only in 30% of hepatocytes. Infected cells presented at their surface epitopes coming from structural proteins, while infected virus-producing cells presented all HCV antigens, structural and non structural.
Because HCV targets a vital organ such as the liver, therapeutic vaccine need to be very specific in order to avoid complete liver destruction and death of the patients. Indeed, we choose for our therapeutic vaccine antigens that are only found in infected virus producing hepatocytes, and then target antigen will be non-structural proteins. Moreover, NS3 and NS4b are highly immunogenic in chronic infected patients, as efficient as structural core or E1 E2 structural proteins. Therefore the present inventors included NS5b as an antigen too.
In one embodiment, NS3 and NS5b were chosen because of their possible expression as recombinant protein and the availability of their 3D structure.
Description of an embodiment of a vaccine: A particular embodiment of a vaccine consisted of bifunctional antibodies, which were directed against Dendritic Cells specific receptors and have target antigens fused at C terminus part of heavy chain. This allows unique targeting of DC and more precisely different DC subset that expressed different receptors, DC activation through the targeted receptor, and direct delivery of antigen to DC. In turn antigens are presented more efficiently and APC function is associated to cytokine secretion that orient T cells activation towards different functions.
Design of domains: It is not readily predictable whether any particular non-structural viral protein will be efficiently expressed as a direct antibody-antigen fusion protein. Commonly, fusion proteins may not be soluble and not be secreted. The present application describes that by using flexible linker modules, fragmenting the antigen coding sequence, and varying the fragment order, efficient secretion of recombinant antibody-antigen vaccines bearing extensive stretches of non-structural proteins can be achieved. The current application describes a first testing of constructs by expression of antibody fused to individual HCV non-structural proteins, then linking those that are expressible as soluble protein to each other to maximize the antigen load. Domains were first designed based on the 3D structure of the corresponding full-length proteins. Domains were design as the minimal structured regions in between unfolded loops. Length of the loops was varied in order to increase expression of corresponding domains. Pymol software was used to visualize 3d structures. The domains that expressed at the C-terminal of the antibody heavy chain are represented by SEQ ID NOs: 7-14.
Multiple combinations of individual domains have been made in order to provide as much HCV antigen as possible. In some embodiments, each single domain is separated from the next by flexible linkers, which can be as small as two amino acids (e.g., AS) but can also be longer, e.g., 3, 4, 5, 6, 7 8, 9, 10, 12, 15, 18, 20, 25 or 30 amino acids long. FIG. 1 shows the summary of all combine constructs. The linkers are found in the assembled sequences, can also be SSVSPTTSVHPTPTSVPPTPTKSSP (SEQ ID NO.: 166); PTSTPADSSTITPTATPTATPTIKG (SEQ ID NO.: 167); TVTPTATATPSAIVTTITPTATTKP (SEQ ID NO.: 168); TNGSITVAATAPTVTPTVNATPSAA (SEQ ID NO.: 169).
In another embodiment, domains were also expressed at the C-terminus part of the light chain, and used in combination with heavy chain fused to multiple HCV domains. This allows the formation of a combine antibody with 3HCV domains fused to the heavy chain and one fused to the light chain. FIG. 2 summarizes the construct obtain after fusion of HCV domains at the C-terminus end of light chain.
Preparation of targeting constructs: Anti human DCIR and CD40 V region form H and L chain were cloned in a IgG4 backbone. Spe I cloning site was introduced at the end of the carboxy terminus to clone in frame antigen sequences. HCV antigens from NS3 and NS5b viral proteins represented as subdomains of these proteins were subcloned as a Spe-Not fragment in Nhe-Not linearized pIRES vector.
HCV-domains were designed based on the 3D-structure of the corresponding full-length proteins (PDB code IJXP for NS3protease, 1HEI for NS3Helicase and 1GX5 for NS5b). 3D-structures were visualized using PyMol software. Domains were designed as the minimal structured regions in between unfolded loops. Length of the loops was varied in order to increase expression of corresponding domains fused to the recombinant antibody. For multiple domains cloning, linkers were introduced between domains using Spe-Not/Nhe-Not strategy. Mosaic sequences, used in this study, corresponding to the maximum HCV-domains expressed as antibody-antigen recombinant fusion proteins are shown below. They included amino acids 95 to 180 from NS3Protease, amino acids 132 to 254 from NS3Helicase and a recombinant fusion of amino acids 55 to 80; 172 to 261 and 276 to 362 from NS5bPolymerase. Spe, Nhe and Not introduced cloning sites are underlined.
SEQ ID NOS: 1-6 show the amino acid sequence of the HCV proteins E1, E2, NS3, and NS5b mosaic sequences. Membrane domains are underlined. The full-length protein NS3 contains 631 amino acids and is also presented as being cut in its two enzymatic activities proteins: NS3Protease and NS3Helicase. These may also be produced as recombinant proteins N-terminal fused to either histidine tag or Cohesin tag.







Envelop protein E1 (192 amino acids)


(SEQ ID NO: 1):


YQVRNSSGLYHVTNDCPNSSIVYEAADAILHTPGCVPCVREGNASRCWVA


VTPTVATRDGKLPTTQLRRHIDLLVGSATLCSALYVGDLCGSVFLVGQLF


TFSPRRHWTTQDCNCSIYPGHITGHRMAWDMMMNWSPTTAVVAQLLRIPQ


AILDMIAGAHWGVLAGIAYFSMVGNWAKVLVVLLLFAGVDA


 


Envelop protein E2 (363 amino acids)


(SEQ ID NO: 2):


ETHVTGGSAARTTAGLAGLFTPGAKQNIQLINTNGSWHINRTALNCNDSL


NTGWVAGLFYYHKFNSSGCPERLASCRPLTDFDQGWGPISYANGSGPDQR


PYCWHYPPKPCGIVPAKSVCGPVYCFTPSPVVVGTTDRSGAPTYNWGEND


TDVFVLNNTRPPLGNWFGCTWMNSTGFTKVCGAPPCVIGGVGNNTLHCPT


DCFRKHPEATYSRCGSGPWITPRCLVDYPYRLWHYPCTINYTIFKIRMYV


GGVEHRLEAACNWTRGERCDLEDRDRSELSPLLLSTTQWQVLPCSFTTLP


ALSTGLIHLHQNIVDVQYLYGVGSSIASWAIKWEYVVLLFLLLADARVC


SCLWMMLLISQAEA


 


Non structural protein 3 NS3 (FL 631 amino acids)


(SEQ ID NO: 3):


APITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQIVSTAAQTFLATCING


VCWTVYHGAGTRTIASPKGPVIQMYTNVDQDLVGWPAPQGARSLTPCTCG


SSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGHA


VGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQSFQVA


HLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDP


NIRTGVRTITTGSPITYSTYGKFLADGGCSGGAYDIIICDECHSTDATSI


LGIGTVLDQAETAGARLVVLATATPPGSVTVPHPNIEEVALSTTGEIPFY


GKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLDVSVI


PTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTT


LPQDAVSRTQRRGRTGRGKPGIYRFVAPGERPSGMFDSSVLCECYDAGCA


WYELTPAETTVRLRAYMNTPGLPVCQDHLEFWEGVFTGLTHIDAHFLSQT


KQSGENLPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPTPLLY


RLGAVQNEVTLTHPITKYIMTCMSADLEVVT


 


NS3 (prot 189 amino acids) (SEQ ID NO: 4):


APITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQIVSTAAQTFLATCING


VCWTVYHGAGTRTIASPKGPVIQMYTNVDQDLVGWPAPQGARSLTPCTCG


SSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGHA


VGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQS


 


NS3 (hel 442 amino acids) (SEQ ID NO: 5):


FQVAHLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAH


GIDPNIRTGVRTITTGSPITYSTYGKFLADGGCSGGAYDIIICDECHSTD


ATSILGIGTVLDQAETAGARLVVLATATPPGSVTVPHPNIEEVALSTTGE


IPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLD


VSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTI


ETTTLPQDAVSRTQRRGRTGRGKPGIYRFVAPGERPSGMFDSSVLCECYD


AGCAWYELTPAETTVRLRAYMNTPGLPVCQDHLEFWEGVFTGLTHIDAHF


LSQTKQSGENLPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPT


PLLYRLGAVQNEVTLTHPITKYIMTCMSADLEVVT


 


Non structural NS5b (591 Amino acids)


(SEQ ID NO: 6):


SMSYSWTGALVTPCAAEEQKLPINALSNSLLRHHNLVYSTTSRSACQRQK


KVTFDRLQVLDSHYQDVLKEVKAAASKVKANLLSVEEACSLTPPHSAKSK


FGYGAKDVRCHARKAVNHINSVWKDLLEDSVTPIDTTIMAKNEVFCVQPE


KGGRKPARLIVFPDLGVRVCEKMALYDVVSKLPLAVMGSSYGFQYSPGQR


VEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDIRTEEAIYQCCDLDPQAR


VAIKSLTERLYVGGPLTNSRGENCGYRRCRASGVLTTSCGNTLTCYIKAR


AACRAAGLQDCTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPP


GDPPQPEYDLELITSCSSNVSVAHDGAGKRVYYLTRDPTTPLARAAWETA


RHTPVNSWLGNIIMFAPTLWARMILMTHFFSVLIARDQLEQALDCEIYGA


CYSIEPLDLPPIIQRLHGLSAFSLHSYSPGEINRVAACLRKLGVPPLRAW


RHRARSVRARLLSRGGRAAICGKYLFNWAVRTKLKLTPIAAAGQLDLSGW


FTAGYSGGDIYHSVSHARPRWFWFCLLLLAAGVGIYLLPNR






The nucleotide sequences are presented herein below.







NS3Protease domain B (SEQ ID NO: 145)


ACTAGTACTCCTTGTACCTGCGGCTCATCCGACCTGTACCTGGTCACCCG


GCACGCAGACGTCATTCCTGTACGCCGACGCGGGGATAGTAGGGGGAGCC


TGCTCTCTCCAAGACCCATATCCTACCTCAAGGGCAGCAGCGGTGGACCA


CTGCTGTGTCCCGCTGGTCATGCTGTGGGAATATTTAGGGCCGCAGTGTG


TACCAGAGGCGTGGCCAAAGCTGTTGATTTTATTCCCGTCGAAAATCTTG


AAACAACCATGAGAAGCCCAGTGTTCACAGACAACTCATCTCCCCCAGCA


GTGCCGCAGAGTGCTAGCTGAGAATTCGCGGCCGC


 


NS3Helicase domain B (SEQ ID NO: 146):


ACTAGTGTGACTGTGCCCCACCCCAATATCGAAGAGGTGGCCCTTAGTAC


TACCGGGGAAATTCCTTTCTACGGGAAGGCCATCCCTCTCGAGGTTATTA


AAGGAGGGCGACATCTGATTTTTTGCCACTCCAAGAAGAAGTGTGACGAG


CTGGCCGCGAAACTGGTTGCCTTGGGCATCAACGCTGTCGCATACTATCG


GGGACTGGATGTATCAGTGATACCCACCAGCGGAGTGGTAGTTGTCGTCG


CTACAGACGCATTGATGACCGGCTTTACAGGAGATTTCGACTCCGTCATC


GACTGTAACACATGCGTGACTCAGACAGTGGATTTCAGCCTTGACCCGAC


GTTTACGATTGAGACCACCACTCTCCCTCAGGATGCTGTGTCTAGGACCC


AAAGACGCGGTCGCACAGGCCGGGGCAAACCAGGCATCTATAGGTTCGTG


GCACCAGGGGAAAGAGCTAGCTGAgaattcGCGGCCGC


 


NS5bPalm (SEQ ID NO: 147):


ACTAGTGTGCTGGACTCTCACTACCAGGATGTCCTGAAGGAAGTAAAAGC


AGCCGCTTCTAAAGTCAAAGCGAACGCTCTGTACGATGTCGTTTCCAAAC


TGCCGCTGGCTGTCATGGGCTCTTCCTACGGCTTTCAGTATTCCCCGGGT


CAGCGCGTTGAGTTCCTGGTCCAGGCGTGGAAATCCAAAAAGACTCCGAT


GGGTTTTTCCTATGACACTCGCTGCTTCGACAGCACCGTTACCGAAAGCG


ACATTCGCACCGAGGAAGCAATCTACCAGTGCTGCGACCTGGACCCACAG


GCCCGCGTGGCGATCAAATCTCTGACCGAACGCCTGTACGTTGGCCGCTG


TCGCGCTTCCGGTGTTCTGACGACCTCCTGCGGTAATACGCTGACCTGCT


ACATCAAAGCACGCGCTGCCTGTCGCGCAGCCGGTCTGCAGGACTGCACC


ATGCTGGTGTGTGGCGATGACCTGGTGGTGATCTGCGAAAGCGCTGGCGT


GCAGGAAGACGCAGCAAGCCTGCGCGCTTTCACCGAAGCTATGACTCGCT


ACTCTGCGCCGCCGGGTGACCCGCCGCAGCCAGAATACGATCTGGAGCTG


ATCACCGCTAGCTAAGAATTCGCGGCCGC






SEQ ID NOS: 7-14 show the HCV antigen domains E1a, E2, ProtA, Prot B, Hel A, Hel B, Hel C, and NS5 bpalm. These were expressed as antibody fusion proteins. For all constructs, amino acids TS and AS (shown in red) have been added for cloning purpose to the mosaic HCV sequence. NS5b palm has been constructed based on NS5b 3D structure (1C2P). It is based on structural domain corresponding of the palm domain of NS5b polymerase and do not correspond to the linear amino acid sequence;







Envelop protein E1a construct (63 amino acids)


(SEQ ID NO: 7):


TSVGQLFTFSPRRHWTTQDCNCSIYPGHITGHRMAWDMMMNWSPTTAVVA


 


QLLRIPQAILDMIAGAS






In SEQ ID NO: 7 membrane domain and predicted unfolded regions have been removed from E1 mosaic 192 aa sequence to increase expression of the Ab fusion protein.







Envelop protein E2 mosaic sequence (342 amino


acids) (SEQ ID NO: 8):


TSETHVTGGSAARTTAGLAGLFTPGAKQNIQLINTNGSWHINRTALNCND


 


SLNTGWVAGLFYYHKFNSSGCPERLASCRPLTDFDQGWGPISYANGSGP


 


DQRPYCWHYPPKPCGIVPAKSVCGPVYCFTPSPVVVGTTDRSGAPTYN


 


WGENDTDVFVLNNTRPPLGNWFGCTWMNSTGFTKVCGAPPCVIGGVGNNT


 


LHCPTDCFRKHPEATYSRCGSGPWITPRCLVDYPYRLWHYPCTINYTIFK


 


IRMYVGGVEHRLEAACNWTRGERCDLEDRDRSELSPLLLSTTQWQVLPCS


 


FTTLPALSTGLIHLHQNIVDVQYLYGVGSSIASWAIKWEYVVLLFLLAS






In SEQ ID NO: 8 the membrane domain has been removed for E2 mosaic sequence.
NS3Protease has been cut in 2 structural domains based on its 3D structure (IJXP).







ProtA


(SEQ ID NO: 9)


TSAPITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQIVSTAAQTFLATCI


NGVCWTVYHGAGTRTIAS


 


Prot B


(SEQ ID NO: 10)


TSTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGP


LLCPAGHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPA


VPQSAS


 


NS3 Helicase enzymatic protein has been cut in 3


structural domains based on NS3 Helicase 3D


structure. (1HEI)


Hel A


(SEQ ID NO: 11)


 FQVAHLHAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSK


AHGIDPNIRTGVRTITTGSPITYSTYGKFLADGGCSGGAYDIIICDECHS


TDATSILGIGTVLDQAETAGARLVVLATATPPGSAS


 


Hel B


(SEQ ID NO: 12)


 VTVPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDE


LAAKLVALGINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVI


DCNTCVTQTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGRGKPGIYRFV


APGERAS


 


Hel C


(SEQ ID NO: 13)


 PSGMFDSSVLCECYDAGCAWYELTPAETTVRLRAYMNTPGLPVCQDHL


EFWEGVFTGLTHIDAHFLSQTKQSGENLPYLVAYQATVCARAQAPPPSWD


QMWKCLIRLKPTLHGPTPLLYRLGAVQNEVTLTHPITKYIMTCMSADLEV


VTAS


 


NS5bpalm


(SEQ ID NO: 14)


TSVLDSHYQDVLKEVKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPG


QRVEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDIRTEEAIYQCCDLDPQ


ARVAIKSLTERLYVGRCRASGVLTTSCGNTLTCYIKARAACRAAGLQDCT


MLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPPGDPPQPEYDLEL


ITAS






HCV sequence and HCV domains constructions: Due to the high polymorphism of HCV, a sequence that is representative of most of circulating HCV sequence was selected.
A mosaic sequence was derived using the mosaic vaccine tools at http://www.hiv.lanl.gov/content/sequence/MOSAIC/ interface choosing mosaic sequence cocktail, 1 as cocktail size and 9 as epitope size. We used 213 sequences for NS3 mosaic and 310 sequences for NS5b mosaic. All sequences correspond to complete genes of 1893, 1773 nucleotides respectively and found in euHCVdb (available on the internet at: euhcvdb.ibcp.fr/euHCVdb/).
Synthetic corresponding genes were purchased from Bio Basic Inc. (Ontario Canada). For cloning purposes, Spe cloning site was introduced at 5′ end and Nhe, EcoRI and Not I at the 3′ end. HCV domains were then constructed by PCR. NS3Protease domain B was construct using the synthetic gene cloned in pUC57 as template and the following primers: NS3Protease domain B forward: 5′-GAGCTCGGATCCACTAGTACTCCTTGTACCTGCGGCTCATCC−3′ (SEQ ID NO: 148) NS3Protease domain B reverse: 5′-GCCCGCGGCCGCGAATTCTCAGCTAGCACTCTGCGGCACTGCTGGGGG-3′ (SEQ ID NO: 149). NS3Helicase domain B was ordered directly as a synthetic gene. For NS5bPolymerase Palm domain construction, regions coding for amino acids 172 to 261 and 276 to 362 were amplified using NS5b synthetic gene and the respective following primers: Ns5b Palm (aa 172-261) forward: 5′-TCTAAAGTCAAAGCGAACGCTCTGTACGATGTCGTTTCC-3′ (SEQ ID NO: 150), Ns5b Palm (aa 172-261) reverse: 5′-ACCGGAAGCGCGACAGCGGCCAACGTACAGGCGTTCGGT-3′ (SEQ ID NO: 151), NS5b Palm (aa 276-362) forward: 5′-ACCGAACGCCTGTACGTTGGCCGCTGTCGCGCTTCCGGT-3′ (SEQ ID NO: 152), NS5b Palm (aa 276-362) reverse: 5′-GCGGCCGCGAATTCttAGCTAGCGGTGATCAGCTCCAG-3′ (SEQ ID NO: 153). Amplified products were then used as templates together with annealed primers 5′-CAAGCCCAACCCCACTAGTGTGCTGGACTCTCACTACCAGGATGTCCTGAAGGAAGTAAAAG CAGCCGCTTCTAAAGTCAAAGCGAACGCTCTGTACGAT-3′ (SEQ ID NO: 154) and 5′-ATCGTACAGAGCGTTCGCTTTGACTTTAGAAGCGGCTGCTTTTACTTCCTTCAGGACATCCTG GTAGTGAGAGTCCAGCACACTAGTGGGGTTGGGCTTG-3′ (SEQ ID NO: 155) in a final PCR using primers 5′-CAAGCCCAACCCC-3′ (SEQ ID NO: 156) and 5′-GCGGCCGCGAATTCTTAGCTAGCGGTGATCAGCTCCAG-3′ (SEQ ID NO: 157). The amplified NS5bPolymerase Palm domain was then cloned in TA vector and sub-cloned in XX vector using Nhe/Not strategy.
Chimeric Recombinant Antibodies Purification: For construct selection, chimeric DC-specific antibodies were transiently expressed in HEK293 cells and purified from the supernatant using Protein A sepharose chromatography. DNA from chimeric constructs expressed in HEK293 was then sub-cloned in cetHSpuro vector as AgeI/NotI fragment for expression in CHO cells after stable transfection. Antibodies were purified from supernatants using ProteinA sepharose.
Patients were recruited at the Baylor Hospital Liver Transplant Clinic (BHLTC, Dallas, Tex.) after obtaining informed consent. The study was approved by the Institutional Review Board of the Baylor Health Care System (Dallas, Tex.). Peripheral blood (100 ml) was collected at the BHLTC from 29 chronic HCV-infected adult patients and one healthy donor in contact with chronic HCV-infected patient. Leukapheresis were collected at Baylor University Medical Center Apharesis Collection Center (Dallas, Tex.) from all the enrolled individuals within 30 days after the first visit. Patient information is summarized in Table I.
Preparation of dendritic cells and PBMCs: PBMCs were isolated from heparinized blood on Ficoll density gradients. Monocytes were enriched from the leukapheresis according to cellular density and size by elutriation (ElutraTM, CaridianBCT, Lakewood, Colo.) as per the manufacturer's recommendations. Elutriation Fraction 5 consisted mainly on monocytes (85% on average). Cells were cryopreserved in 10% DMSO 50% FCS 10% culture medium before use. For dendritic cell generation, monocytes were resuspended in serum-free CellGro DC culture medium (CellGenix Technologie Transfer Gmbh, Germany) at a concentration of 1 106 cells/ml. Media were supplemented with 100 ng/ml granulocyte-macrophage colony-stimulated factor (GMCSF, Leukine, Berlex, Wayne, N.J.) and 500 UI/ml alpha-interferon (IFN-α, Intro A, IFN-α-2b, Merck/Schering-Plough, Kenilworth, N.J.). After 24 h of culture at 37 degree Celsius, 5% CO2, fresh cytokines were added. On day 3, recombinant antibody vaccines were added at various concentration (5 nM, 0.5 nM or 0.05 nM) or peptide cluster controls (2 μM each peptide) as indicated. Alternatively, TLR agonists (polyIC, 25 μg/ml; CL075 1 μg/ml; or PAM3, 200 ng/ml; all from Invivogen) were added in the culture at the same time as vaccine candidates or peptide controls. DC were pulsed for 16 h before harvest and used in PBMCs co-culture.







TABLE I







Demographics of patients used in the study.




















Geno-



HCV

Viral


HLA
HLA
HLA


Patient ID
type
Sex
Ethnicity
Race
status
Age
Load
HLA A*
HLA B*
Cw*
DRB1
DQB1





HCV-VAC-001
1a
M
Hispanic
White
non
39
5 877033
0201;
1302;
0202;
0701;
0202;







responder

(H)
0205
5101
0602
1301
0603


HCV-VAC-002
1a
F
Non
White
cured
57
UnDectable
0101;
0818;
0701G;
1101;
0301;





Hispanic

after

(UD)
0301
5108
1502
1301
0603







therapy









HCV-VAC-003
1a
M
Non
White
cured
59
UD
0301;
0702;
0401;
0101;
0501;





Hispanic

after


3004
3501
0702
0402
0302







therapy









HCV-VAC-004
1a
M
Non
White
cured
55
UD
0201;
0702;
0202;
0401;
0302;





Hispanic

after


3201
1002
0702
0901
0303







therapy









HCV-VAC-005
1a
M
Non
White
cured
58
UD
0101;
1801;
0501;
0301;
0503;





Hispanic

after


1101
5101
1402
1407
0201







therapy









HCV-VAC-006
3a
M
Non
White
cured
57
UD
0101;
0702;
0602;
0701;
0303;





Hispanic

after


2902
5701
0702









therapy









HCV-VAC-007
3a
M
Non
White
cured
48
UD
no
no
no
no
no





Hispanic

after


apheresis
apheresis
apheresis
apheresis
apheresis







therapy









HCV-VAC-008
1b
M
Non
White
non
63

0101;
0801;
0701;
0301;
0201;





Hispanic

responder


6901
3508
1203




HCV-VAC-009
1a
M
Non
White
non
51

0101;
0801;
0701G;
0301;
0201;





Hispanic

responder


3004


0701
0303


HCV-VAC-010
1a
M
Non
White
cured
48
UD
0201;
0801;
0202;
0301;
0201;





Hispanic

after


2402
4002
0701G
0701
0202







therapy









HCV-VAC-011
1a
F
Non
White
non
52
?
0205;
1530;
0102;
0301;
0201;





Hispanic

responder


3101
4901
0707G
0802
0402


HCV-VAC-012
1a
M
Non
White
cured
43
UD
0101;
4101;
1502;
0301;
0201;





Hispanic

after


1101
5101
1710G
1305
0301







therapy









HCV-VAC-013
1a
M
Non
White
non
55

0101;
0801;
0401;
0301;
0502;





Hispanic

responder


0201
3502
0701G
1601
0201


HCV-VAC-014
1b
F
Non
White
non
56

3101;
0702;
0401;
0403;
0302;





Hispanic

responder


6801
3503
0702
1501
0602


HCV-VAC-015
2b
M
Non
White
positive
50

0101;
0801;
0401;
0301;
0201;





Hispanic

untreated


0301
3501
0701G
0701
0303


HCV-VAC-016
1a
M
Non
White
non
55

0101;
5001;
0303;
0701;
0503;





Hispanic

responder



5501
0602
1401
0202


HCV-VAC-017
1a
M
Non
White
positive
52

2402;
3901;
0602;
0701;
0202;





Hispanic

untreated


2501
5701
1203

0303


HCV-VAC-018
1a
M
Non
White
non
53

no
no
no
no
no





Hispanic

responder


apheresis
apheresis
apheresis
apheresis
apheresis


HCV-VAC-019
1a
M
Non
White
cured
62
UD
2402;
3901;
0102;
0701;
0202;





Hispanic

after


2501
4402
1203
1501
0602







therapy









HCV-VAC-020
1a
F
Non
White
cured
46
UD
0201;
1501;
0202;
0401;
0301;





Hispanic

after


0301
2705
0304
1101
0302







therapy









HCV-VAC-021
1a
M
Non
White
non
64

0101;
0801;
0304;
0101;
0501;





Hispanic

responder


3002
4001
0701G
0301
0201


HCV-VAC-022
1b
F
Non
White
non
58

0301;
1801;
0304;
0301;
0201;





Hispanic

responder


6801
4001
0501
1302
0604


HCV-VAC-023
2
F
Non
White
positive
45

no
no
no
no
no





Hispanic

untreated


apheresis
apheresis
apheresis
apheresis
apheresis


HCV-VAC-024
2b
M
Non
White
positive
43

0301;
1402;


0202;





Hispanic

untreated



4701


0602


HCV-VAC-025
3a
F
Non
White
positive
31
?
no
no
no
no
no





Hispanic

untreated


apheresis
apheresis
apheresis
apheresis
apheresis


HCV-VAC-026
3a
M
Non
White
positive
29
?
no
no
no
no
no





Hispanic

untreated


apheresis
apheresis
apheresis
apheresis
apheresis


HCV-VAC-027
1a
F
Non
White
positive
26

02; 26
15(62);


03(7);





Hispanic

untreated



51


05


HCV-VAC-028
1a
F
Hispanic
other
positive
47

03; 25
07; 18


06;







untreated









HCV-VAC-029
Non
F
Non
White
uninfected
63

01; 11
44; 55


02; 05



infected

Hispanic











HCV-VAC-030
1
M
Non
White
positive
57
?
03; 24
07; 27


03(7);





Hispanic

untreated






06









Expansion of Antigen-specific T cells in DC/PBMCs coculture. Frozen PBMCs from leukapharesis were thawed, washed by centrifugation and resuspended at 2×106 cells/ml in cRPMI medium. Autologous DC loaded with vaccine candidates or peptides cluster controls were co-cultured with PBMCs in a 24 well tissue plate at a ratio of 1/20 and incubated for a total of 10 days. IL2 (20 IU/ml, Aldesleukine, ProleukineR; Bayer Healthcare and Novartis, Emeryville, Calif.) was added every two days. At day 9, PBMCs from a 24-well plate were washed, distributed in 2 wells in a 96-well plates and rested for 24 h. The specificity of the T-cell response elicited by vaccine candidate loaded-DC was assessed by restimulation of PBMCs with peptide clusters (2 μM each peptide). For each condition, a negative background control was included as a restimulation without peptides.
Flow cytometry: After 1 hour of peptide clusters restimulation, BFA (Sigma) was added for the last 5-6 h to block cytokine secretion. The cells were stained for surface markers with a combination of fluorochrome antibodies (perCP-CD3, PE-CD8, APCH7-CD4), fixed, permeabilized and intracellular-stained with a mixture of APC-IFNγ, FITC-IL2 and PEcy7-TNFα antibodies. For CTL marker function analysis, FITC-CD107a antibody was added with BFA in the culture medium and the following antibodies combination was used for the surface staining: PerCP-CD3, pacific blue-CD8, APCH7-CD4 and for the intracellular staining: PE-IFNγ, APC-GranzB, APCcy7-TNFα. All antibodies were purchased from BD sciences except APC-GranzB (Invitrogen). Cells were analyzed on a FACS-Canto collecting 500,000 events, and results analyzed using FlowJo software. Most of the data were displayed as two colors plot to measure IFN-γ and TNF-α production in CD3+CD8+ or CD3+CD4+ cells.
Luminex: Supernatants of DC-PBMCs co-culture were harvested 48 h after PBMCs restimulation with peptide clusters. Cytokine multiplex assays were employed to analyzed IFN-γ, IL-10, and IL-13.
Evaluation of embodiments of vaccines: Vaccine candidate were tested in targeting experiment by co-culture of vaccine with PBMCs from chronic HCV infected patients or chronic HCV infected patients cured after IFNa-Ribavirin therapy. The data show that anti-CD40 or anti-DCIR vaccines bearing a HCV NS3HelB antigen can recall a potent memory antigen-specific anti-CD4+ T cell response in vitro using immune cells from HCV infected patients. In this in vitro culture system anti-CD40 and anti-DCIR are equally potent vaccines—these DCs express both receptors. Anti-DCIR vaccine construct bearing longer HCV antigen coverage induced multifunctional CD4+ antigen specific T cells against multiple HCV epitopes.
The data further show that anti-DCIR vaccines bearing a HCV NS3HelBC antigen can recall a potent memory antigen-specific anti-CD4+ T cell response in vitro using immune cells from HCV infected patients. This response is directed against multiples HCV epitopes. In this in vitro culture system, both concentration used for anti-DCIR HCV-NS3HelBC targeting are equally potent in contrast to anti-DCIR HCV-NS3HelB vaccine.
FIGS. 3A-3B demonstrate the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB specific antigen to elicit the expansion of antigen-specific CD4+ T cells from a chronic HCV infected patient cured after IFNa-Ribavirin therapy. Delivering NS3HelB to DCs through CD40 and DCIR induces IFNγ-TNFa-producing HCV NS3HelB-specific CD4+ T cells. PBMC cells from chronic HCV infected patients; either cured after therapy or in treatment failure, were co-cultured with IFNDCs targeted with anti-CD40-NS3HelB or anti-DCIR-NS3HelB for 10 days. Cells were stimulated with peptides clusters (10 peptides of 15-mers in each clusters) covering HCV NS3 HelB (10 μM): FIG. 3A after 2 days, culture supernatants were analyzed for measuring IFNγ and FIG. 3B PBMC cells were stained for measuring the frequency of peptide-specific CD4+ T cells intracellular IFNγ+TNFα+ cells.
Longer construct are equally potent to recall multi epitopes HCV specific T cells. The data in e.g., FIG. 4, show that both anti-CD40 and anti-DCIR vaccines bearing HCV NS3HelB NS3ProtB and NS5BPalm antigens can recall a potent memory antigen-specific anti-CD4+ T cell response in vitro using immune cells from HCV infected patients cured after IFN-Ribavirin therapy. This response is directed against multiples HCV epitopes. In this in vitro culture system, dose effect is observed consistent with clear targeting, with an optimum concentration being at 5 nM for anti-CD40 construct and 0.5 nM for anti-DCIR construct. At these concentrations IgG4 controls induce significantly lower CD4+ T cells responses, consistent with antibody targeting of DC.
FIGS. 5A to 5C demonstrate the ability of recombinant anti-DCIR and anti-CD40 antibodies fused to HCV NS3HelB, HCV NS3ProtB and HCV NS5BPalm specific antigens to elicit the expansion of antigen-specific CD4+ T cells from a chronic HCV infected patient cured after IFNa-Ribavirin therapy. Delivering HCV antigen to DCs through CD40 and DCIR induces IFNγ-TNFa-producing HCV-specific CD4+ T cells, with multi epitopes, specific CD4 T cells. PBMC cells from chronic HCV infected patients cured after therapy were co-cultured with autologous IFNαDCs targeted with anti-CD40-NS3HelB-NS3ProtB-NS5BPalm or anti-DCIR-NS3HelB-NS3ProtB-NS5BPalm for 10 days. Cells were stimulated with peptides clusters (10 peptides of 15-mers in each clusters) covering HCV NS3HelB, NS3ProtB or NS5BPalm (2 μM). PBMC cells were stained for measuring the frequency of peptide-specific CD4+ T cells intracellular IFNγ+TNFα+ cells.
The data in FIGS. 6A to 6C demonstrate that anti-CD40 vaccines bearing HCV NS3HelB NS3ProtB and NS5BPalm antigens can recall a potent memory antigen-specific anti-CD8+ T cell response in vitro using immune cells from HCV infected patients cured after IFN-Ribavirin therapy. This response is directed against multiples HCV epitopes. In this in vitro culture system, dose effect is observed consistent with clear targeting, of DC with an optimum concentration being at 5 nM for anti-CD40 constructs. At these concentrations IgG4 controls induce significantly no CD8+ T cells responses, consistent with antibody targeting of DC.
Similar responses are induced in multiple different chronic HCV infected patients either cured or after therapy or in treatment failure.
The data in FIGS. 7A to 7D show that all chronic HCV infected patients cured after therapy are able to recall CD4+ T cells memory after co-culture of PBMCs with DC targeted with either anti-CD40 or anti-DCIR or both, construct bearing HCV antigens.
The data in FIGS. 8A to 8D shows that all chronic HCV infected patients in treatment failure are able to recall CD4+ T cells memory after co-culture of PBMCs with DC targeted with either anti-CD40 or anti-DCIR or both, construct bearing HCV antigens. Compare to chronic HCV infected patients cured after therapy, responses are low in chronic HCV infected patients in treatment failure and more antigen dependent since for example HCV-VAC-016 patient has no CD4+ T cells memory cells recalled after DC targeting with NS5bPalm construct.
CD8+ antigen specific T cells were obtained after TLR agonist introduction in the co-culture of vaccine with PBMC cells from HCV patients.
The data in FIGS. 9A and 9B show that TLR2 triggering with PAM3 during DC targeting with anti-DCIR vaccines bearing a HCV NS3HelB antigen can recall a potent memory antigen-specific anti-CD4+ and CD8+ T cell response in vitro using immune cells from HCV infected patients. Moderated CD8+ response is also induced by TLR3 triggering and no CD4+ or CD8+ response is induced after TLR7/8 triggering by CL097 in this study. Similar responses are induced in multiple different chronic HCV infected patients either cured or after therapy or in treatment failure.
The data in FIGS. 10A-10D show that TLR2 triggering with PAM3 during DC targeting with anti-CD40 or anti-DCIR vaccines bearing a HCV NS3HelB or HCV NS3ProtB antigen can recall a potent memory antigen-specific anti-CD4+ and CD8+ T cell responses in vitro using immune cells from HCV infected patients. Moderated CD8+ response is also induced by TLR3 triggering in some patients, and cyclic glucan can dramatically increase CD8+ T cells responses in one patient.
FIG. 11 demonstrates the ability of combination of TLR agonists and anti-CD40 HCV-constructs to increase CD4+ T cells responses in chronic HCV infected patients in treatment failure. HCV antigens from NS3 Helicase HelB or from NS3 Protease ProtB constructs were delivered to DCs through CD40 or DCIR. IFNαDCs were targeted with anti-CD40-NS3HelB, anti-DCIR-NS3HelB, anti-CD40-NS3ProtB, anti-DCIR-NS3ProtB, in presence of PAM3 (TLR2 agonist; 200 ng/ml), CL095 (TLR7/8 agonist; 5 μg/ml) or polyIC (TLR3 agonist; 25 μg/ml) or cyclic glucan (TLR4 agonist, 10 μg/ml) before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB constructs or with peptide clusters C3 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB constructs. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS.
All tested HCV patients are able to recall CD4+ and CD8+ HCV specific memory after DC-targeting with HCV vaccine candidates.
FIG. 12A to 12C demonstrate the ability of HCV vaccine candidates to recall CD4+ T cells responses in all chronic HCV infected patients (cured or in treatment failure). HCV antigens from NS3 Helicase HelB, NS5b polymerase Palm or from NS3 Protease ProtB constructs were delivered to DCs through CD40 (FIG. 12B) or DCIR (FIG. 12C). IFNaDCs were targeted with anti-CD40-[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain], anti-DCIR-[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain] before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7-C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB domain, with peptide clusters C2-C3-C4 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB domain or with peptide clusters C2-C4-C5-C6-C7 (10 μM; 10 peptides of 15-mers) covering HCV NS5b Palm domain. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ and CD8+ intracellular IFNγ+TNFa+ cells, and analyzed by FACS. The number of CD4+ IFNg+TNFa+ cells induced vaccine candidate is shown.
It was also observed that different combinations of HCV domains on vaccine candidate are equally equivalent to recall CD4+HCV memory. Moreover, HCV antigen combination where two domains are borne on heavy chain and one on light chain is more efficient than having the 3 borne by heavy chain.
FIGS. 13A-13E demonstrate the ability of different HCV antigen combination on vaccine candidate for recall CD4+ T cells responses in chronic HCV infected cured patients. HCV antigens from NS3 Helicase HelB, NS5b polymerase Palm or from NS3 Protease ProtB combination constructs were delivered to DCs through CD40 or DCIR. IFNαDCs were targeted with second-generation vaccines anti-CD40-[NS3HelB on light chain and NS3ProtB ̃NS5bPalm on heavy chain], anti-DCIR-[NS3HelB on light chain and NS3ProtB ̃NS5bPalm on heavy chain], or first-generation vaccines anti-CD40-[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain], anti-DCIR—[NS3HelB ̃NS3ProtB ̃NS5bPalm on heavy chain] before co-culture for 10 days with PBMC cells from chronic HCV infected patients cured after therapy. Cells were stimulated for 6 h with peptide clusters C7 and C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB domain (shown in green on the figure), with peptide clusters C2-C3-C4 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB domain (shown in pink on the figure) or with peptide clusters C2-C4-C5-C6-C7 (10 μM; 10 peptides of 15-mers) covering HCV NS5b Palm domain (shown in orange in the figure). PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS. The number of CD4+IFNγ+TNFα+ cells induced by first-generation vaccine or second-generation vaccine is compared in the last panel.
The vaccine candidates described in the present invention also showed the ability induce cross reactivity recall memory responses in patients infected with an HCV genotype different from those used to build the vaccine (FIGS. 14A to 14H). FIGS. 14A to 14H demonstrate ability of vaccine candidate to recall CD4+ T cells responses in HCV patients infected with non 1 genotype and HCV-exposed but non infected individual. HCV antigens from NS3 Helicase HelB, NS5b polymerase Palm or from NS3 Protease ProtB combination constructs were delivered to DCs through CD40 or DCIR and DC loaded were co-culture for 10 days with PBMC cells from HCV patients infected with non 1 genotype HCV-infected patients (HCV-015, 2b) and HCV-exposed but non infected individual (HCV-029). Cells were stimulated for 6 h with peptide clusters C7 and C9 (10 μM; 10 peptides of 15-mers) covering HCV NS3 HelB domain, with peptide clusters C2-C3-C4 (10 μM; 10 peptides of 15-mers) covering HCV NS3 ProtB domain or with peptide clusters C2-C4-C5-C6-C7 (10 μM; 10 peptides of 15-mers) covering HCV NS5b Palm domain. PBMC cells were stained for measuring the frequency of peptide-specific CD4+ intracellular IFNγ+TNFα+ cells, an analyzed by FACS.
FIGS. 15A and 15B show the results from a 10 day expansion culture whereby a dose range of 1st generation anti-DCIR-HCV vaccine (left panels) is compared to second generation anti-DCIR-HCV vaccine (right panels). Doses were 0.05 nM, 0.5 nM, and 5 nM and antigen-specific responses were ascertained by stimulation with no peptide (control) or ProtA, HelB, or Palm peptide pools in the presence of Brefeldin, followed by staining for CD3+, CD4+ and intracellular IFNg and TNFa. Samples were analyzed by FACS. Shown are comparable CD4+ HCV antigen-specific responses to the two generations of vaccines.
FIGS. 16A and 16B show the results from a 10 day expansion culture whereby a dose range of 1st generation anti-CD40-HCV vaccine (left panels) is compared to second generation anti-CD40-HCV vaccine (right panels). Doses were 0.05 nM, 0.5 nM, and 5 nM and antigen-specific responses were ascertained by stimulation with no peptide (control) or ProtA, HelB, or Palm peptide pools in the presence of Brefeldin, followed by staining for CD3+, CD4+ and intracellular IFNg and TNFa. Samples were analyzed by FACS. Shown are comparable CD4+ HCV antigen-specific responses to the two generations of vaccines.
Non-limiting examples different DC-specific antibodies or fragments (both nucleotide and protein sequences) that may be used in the preparation of the HCV vaccine of the present invention are shown herein below, the nomenclature corresponding to the target (e.g., Anti_CLEC_6_9B9.2G12_Heavy Hv-V-hIgG4H-C—is an anti-CLEC-6 antibody from the mouse hybridoma clone “9B9.2G12” (which is the source of the anti-CLEC-6 antibody sequence); heavy chain “H” variable region “v” (which can be humanized) heavy and is an IgG4 constant region isotype. The same nomenclature applies to light chains (from mouse Kappa light chains), and the antigens.







Anti_CLEC_6_9B9.2G12_Hv-V-hIgG4H-C


(SEQ ID NO: 15):


ATGGGCAGGCTTACTTCTTCATTCTTGCTACTGATTGTCCCTGCATATGT


CCTGTCCCAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCT


CCCAGACCCTCAGTCTGACCTGTTCTTTCTCTGGGTTTTCACTGAGCACT


TCTGGTATGAGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGA


GTGGCTGGCTCACATTTGGTGGAATGATGATAAGTACTATAATCCAGTCC


TGAAAAGCCGGCTCACAATCTCCAAGGAGACCTCCAACAACCAGGTATTC


CTCAAGATCGCCAGTGTGGTCTCTGCAGATACTGCCACATACTACTGTGC


TCGATTCTATGGTAACTGTCTTGACTACTGGGGCCAAGGCACCACTCTCA


CAGTCTCCTCGGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCC


TGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAA


GGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGA


CCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTAC


TCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGAC


CTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGA


GAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAG


TTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACAC


TCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGA


GCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAG


GTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA


CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCA


AGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAG


AAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACAC


CCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCT


GCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC


AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTC


CGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGT


GGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC


AACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTG


ATTAATTAA


 


Anti_CLEC_6_9B9.2G12_Heavy (H)v-V-hIgG4H-C


(SEQ ID NO: 80):


MGRLTSSFLLLIVPAYVLSQVTLKESGPGILQPSQTLSLTCSFSGFSLST


SGMSVGWIRQPSGKGLEWLAHIWWNDDKYYNPVLKSRLTISKETSNNQVF


LKIASVVSADTATYYCARFYGNCLDYWGQGTTLTVSSAKTKGPSVFPLAP


CSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY


SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPE


FEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE


VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE


KTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWES


NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH


NHYTQKSLSLSLGKAS


 


Anti_CLEC_6_9B9.2G12_Kv-V-hIgGK-C (SEQ ID NO: 16):


ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGG


TACCAGATGTGATATCCAGATGACACAGACTACATCCTCCCTGTCTGCCT


CTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGC


AATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCT


GATCTACTACACATCAATATTACAATTAGGAGTCCCATCAAGATTCAGTG


GCAGTGGGTCTGAAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAA


GAAGATATTGCCACTTACTTTTGCCAACAGGGTGATTCGCTTCCATTCAC


GTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti_CLEC_6_9B9.2G12_Light (K)v-V-hIgGK-C


(SEQ ID NO: 81):


MMSSAQFLGLLLLCFQGTRCDIQMTQTTSSLSASLGDRVTISCRASQDIS


NYLNWYQQKPDGTVKLLIYYTSILQLGVPSRFSGSGSETDYSLTISNLEQ


EDIATYFCQQGDSLPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-ASGPR_49C11_7H-LV-hIgG4H-C (SEQ ID NO: 17):


ATGAGAGCGCTGATTCTTTTGTGCCTGTTCACAGCCTTTCCTGGTATCCT


GTCTGATGTGCAGCTTCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTC


AGTCACTTTCACTCACCTGCACTGTCACTGGCTACTCCATCACCAGTGGT


TATAGCTGGCACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAATGGAT


GGGCTACATACTCTTCAGTGGTAGCACTAACTACAACCCATCTCTGAAAA


GTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAG


TTGAATTCTGTGACTACTGAGGACACAGCCACATATTTCTGTGCAAGATC


TAACTATGGTTCCTTTGCTTCCTGGGGCCAAGGGACTCTGGTCACTGTCT


CTGCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCC


AGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTA


CTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCG


GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTC


AGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACAC


CTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTG


AGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAA


GGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCAT


GATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGG


AAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCAT


AATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT


GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGT


ACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACC


ATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCC


CCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGG


TCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG


CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG


CTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGG


AGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC


TACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGATTAAT


TAA


 


Anti-ASGPR_49C11_7H-LV-hIgG4H-C (SEQ ID NO: 82):


MRALILLCLFTAFPGILSDVQLQESGPDLVKPSQSLSLTCTVTGYSITSG


YSWHWIRQFPGNKLEWMGYILFSGSTNYNPSLKSRISITRDTSKNQFFLQ


LNSVTTEDTATYFCARSNYGSFASWGQGTLVTVSAAKTTGPSVFPLAPCS


RSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL


SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFE


GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVH


NAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT


ISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNG


QPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNH


YTQKSLSLSLGKAS


 


Anti-ASGPR_49C11_7K-LV-hIgGK-C (SEQ ID NO: 18):


ATGGATTTTCAAGTGCAGATTTTCAGCTTCCTGCTAATCAGTGCCTCAGT


CATAATATCCAGAGGACAAATTGTTCTCACCCAGTCTCCAGCAATCATGT


CTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGCTCAAGT


GTAAGTCACATGCACTGGTACCAGCAGAAGTCAGGCACTTCCCCCAAAAG


ATGGATTTATGACACATCCAGACTGGCTTCTGGAGTCCCTGCTCGCTTCA


GTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAG


GCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTCACCCATG


GTCGTTCGGTGGAGGCACCAAACTCGAGATCAAACGAACTGTGGCTGCAC


CATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT


GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT


ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTG


TCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTG


ACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGT


CACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG


AGTGTTAG


 


Anti-ASGPR_49C11_7K-LV-hIgGK-C (SEQ ID NO: 83):


MDFQVQIFSFLLISASVIISRGQIVLTQSPAIMSASPGEKVTMTCSASSS


VSHMHWYQQKSGTSPKRWIYDTSRLASGVPARFSGSGSGTSYSLTISSME


AEDAATYYCQQWSSHPWSFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGT


ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL


TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-ASGPR_4G2.2_Hv-V-hIgG4H-C (SEQ ID NO: 19):


ATGGCTTGGGTGTGGACCTTGCTATTCCTGATGGCAGCTGCCCAAAGTGC


CCAAGCACAGATCCAGTTGGTGCAGTCTGGACCTGAGCTGAAGAAGCCTG


GAGAGACAGTCAAGATCTCCTGCAAGGCTTCTGGGTATACCTTCACAAAC


TATGGAATGAACTGGGTGAAGCAGGTTCCAGGAAAAGGTTTAAGGTGGAT


GGGCTGGATGGACACCTTCACTGGAGAGCCAACATATGCTGATGACTTCA


AGGGACGGTTTGCCTTCTCTTTGGAAACCTCTGCCAGCACTGCCTATTTG


CAGATCAACAGCCTCAAAAATGAGGACACGGCTACTTATTTCTGTGCAAG


AGGGGGGATTTTACGACTCAACTACTTTGACTACTGGGGCCAAGGCACCA


CTCTCACAGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTG


GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCT


GGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCG


CCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGA


CTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCAC


GAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGG


ACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCA


CCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAA


GGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGG


ACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGC


GTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAG


CACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA


ACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCC


ATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGT


GTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCC


TGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGG


GAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCT


GGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGA


GCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCT


CTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC


TAGCTGATTAATTAA


 


Anti-ASGPR_4G2.2_Hv-V-hIgG4H-C (SEQ ID NO: 84):


MAWVWTLLFLMAAAQSAQAQIQLVQSGPELKKPGETVKISCKASGYTFTN


YGMNWVKQVPGKGLRWMGWMDTFTGEPTYADDFKGRFAFSLETSASTAYL


QINSLKNEDTATYFCARGGILRLNYFDYWGQGTTLTVSSAKTKGPSVFPL


APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG


LYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPA


PEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDG


VEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSS


IEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW


ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA


LHNHYTQKSLSLSLGKAS


 


Anti-ASGPR_4G2.2_Kv-V-hIgGK-C (SEQ ID NO: 20):


ATGAAGTTTCCTTCTCAACTTCTGCTCTTACTGCTGTTTGGAATCCCAGG


CATGATATGTGACATCCAGATGACACAATCTTCATCCTCCTTTTCTGTAT


CTCTAGGAGACAGAGTCACCATTACTTGCAAGGCAAGTGAGGACATATAT


AATCGGTTAGGCTGGTATCAGCAGAAACCAGGAAATGCTCCTAGGCTCTT


AATATCTGGTGCAACCAGTTTGGAAACTGGGGTTCCTTCAAGATTCAGTG


GCAGTGGATCTGGAAAGGATTACGCTCTCAGCATTACCAGTCTTCAGACT


GAAGATCTTGCTACTTATTACTGTCAACAGTGTTGGACTTCTCCGTACAC


GTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-ASGPR_4G2.2_Kv-V-hIgGK-C (SEQ ID NO: 85):


MKFPSQLLLLLLFGIPGMICDIQMTQSSSSFSVSLGDRVTITCKASEDIY


NRLGWYQQKPGNAPRLLISGATSLETGVPSRFSGSGSGKDYALSITSLQT


EDLATYYCQQCWTSPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-ASGPR_5F10H-LV-hIgG4H-C (SEQ ID NO: 21):


ATGGGATGGAGCTGGATCTTTCTCTTTCTCTTGTCAGGAACTGGAGGTGT


CCTCTCTGAGGTCCAGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTG


GGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACCTTCACTGAC


TACTACATGAAGTGGGTGAAGCAGAGCCATGGAAAGAGCCTTGAGTGGAT


TGGAGATATTAATCCTAACTATGGTGATACTTTCTACAACCAGAAGTTCG


AGGGCAAGGCCACATTGACTGTAGACAAATCCTCCAGGACAGCCTACATG


CAGCTCAACAGCCTGACATCTGAGGACTCTGCAGTCTATTATTGTGGAAG


AGGGGACTATGGATACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCG


TCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGC


TCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGA


CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCA


GCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCC


CTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTA


CACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAG


TTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTC


GAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCT


CATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCC


AGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG


CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCG


TGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGG


AGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAA


ACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCT


GCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC


TGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT


GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGA


CGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGC


AGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC


CACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-ASGPR_5F10H-LV-hIgG4H-C (SEQ ID NO: 86):


MGWSWIFLFLLSGTGGVLSEVQLQQSGPELVKPGASVKMSCKASGYTFTD


YYMKWVKQSHGKSLEWIGDINPNYGDTFYNQKFEGKATLTVDKSSRTAYM


QLNSLTSEDSAVYYCGRGDYGYFDVWGAGTTVTVSSAKTKGPSVFPLAPC


SRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS


LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEF


EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV


HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK


TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN


GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN


HYTQKSLLSLGKAS


 


Anti-ASGPR_5F10K-LV-hIgGK-C (SEQ ID NO: 22):


ATGGAGACACATTCTCAGGTCTTTGTATACATGTTGCTGTGGTTGTCTGG


TGTTGAAGGAGACATTGTGATGACCCAGTCTCACAAATTCATGTCCACAT


CAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGGATGTGGGT


ACTGCTGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCTAAACTACT


GATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCTTCACAG


GCAGTGGATCTGGGACAGATTTCACTCTCACCATTAACAATGTGCAGTCT


GAAGACTTGGCAGATTATTTCTGTCAGCAATATAGCAGCAATCCGTACAT


GTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-ASGPR_5F10K-LV-hIgGK-C (SEQ ID NO: 87):


METHSQVFVYMLLWLSGVEGDIVMTQSHKFMSTSVGDRVSITCKASQDVG


TAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTINNVQS


EDLADYFCQQYSSNPYMFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-ASGPR1H11_H-V-hIgG4H-C (SEQ ID NO: 23):


ATGGGATGGAGCTGGATCTTTCTCTTTCTCCTGTCAGGAACTGCAGGTGT


CCTCTCTGAGGTCCAGCTGCAACAGTCTGGACCTGAGTTGGTGAAGCCTG


GGGCTTCAGTGAAGATATCCTGCAAGACTTCTGGATACACATTCACTGAA


TACACCATGCACTGGGTGAGGCAGAGCCATGGAAAGAGCCTTGAGTGGAT


TGGAGGTATTAATCCTATCAATGGTGGTCCTACCTACAACCAGAAGTTCA


AGGGCAAGGCCACATTGACTGTTGACAAGTCCTCCAGCACAGCCTACATG


GAGCTCCGCAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


ATGGGACTATGGTAGTCGAGATGTTATGGACTACTGGGGTCAAGGAACCT


CAGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTG


GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCT


GGTCAAGGACTACTTCCCCGAACCGGTACCGGTGACGGTGTCGTGGAACT


CAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCC


TCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTT


GGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCA


AGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGC


CCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAA


ACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGG


TGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTG


GATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTT


CAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACT


GGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCG


TCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCC


ACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGG


TCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTG


GAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCC


CGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGG


ACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCAT


GAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGG


TAAAGCTAGCTGA


 


Anti-ASGPR1H11_H-V-hIgG4H-C (SEQ ID NO: 88):


MGWSWIFLFLLSGTAGVLSEVQLQQSGPELVKPGASVKISCKTSGYTFTE


YTMHWVRSHGKSLEWIGGINPINGGPTYNQKFKGKATLTVDKSSSTAYME


LRSLTSEDSAVYYCARWDYGSRDVMDYWGQGTSVTVSSAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVPVTVSWNSGALTSGVHTFPAVLQSS


GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP


APEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD


GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE


WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE


ALHNHYTQKSLSLSLGKAS


 


Anti-ASGPR1H11K-LV-var2-hIgGK-C (SEQ ID NO: 24):


ATGGAATCACAGACTCTGGTCTTCATATCCATACTGCTCTGGTTATATGG


TGCTGATGGGAACATTGTAATGACTCAATCTCCCAAATCCATGTCCATGT


CAGTAGGGGAGAGGGTCACCTTGAGCTGCAAGGCCAGTGAGAATGTGGGA


ACTTATGTATCCTGGTATCAACAGAGACCAGAACAGTCTCCAAAACTGCT


GATATACGGGGCATCCAACCGGTACACTGGGGTCCCCGATCGCTTCACAG


GCAGTGGATCTGCAACAGATTTCACTCTGACCATCAGCAGTGTGCAGGCT


GAGGACCTTGCAGATTATCACTGTGGACAGACTTACAGCTATATATTCAC


GTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-ASGPR1H11K-LV-var2-hIgGK-C (SEQ ID NO: 89):


METHSQVFVYMLLWLSGVEGNIVMTQSPKSMSMSVGERVTLSCKASENVG


TYVSWYQQRPEQSPKLLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQA


EDLADYHCGQTYSYIFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-CD1d_2B5.3G10_H-V-hIgG4H-C (SEQ ID NO: 25):


ATGGGATGGAGCCGGATCTTTCTCTTCCTCCTGTCAATAACTGCAGGTGT


CCATTGCCAGGTCCAGGTGCAGCAGTCGGGACCTGAGTTGGTGAAGCCTG


GGGCCTCAGTGAAGATTTCCTGCAAAGCCTCTGGCGACGCATTCAGTAGT


TCTTGGATGAACTGGGTGAAGCAGAGGCCTGGACAGGGTCTTGAGTGGAT


TGGACGGATTTATCTTGGAGATGGAGATATTAATTACAATGGGAAGTTCA


AGGGCAGGGCCACACTGACTGCAGACAAATCCTCCAGCACAGCCTACATG


CAGCTCAGCAGCCTGACCTCTGTGGACTCTGCGGTCTATTTCTGCGCGAG


GCAGCTCGGGCTATGGTATGTTATGGACTACTGGGGTCAAGGAACCTCAG


TCACCGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTGA


 


Anti-CD1d_2B5.3G10_H-V-hIgG4H-C (SEQ ID NO: 90):


MGWSRIFLFLLSITAGVHCQVQVQQSGPELVKPGASVKISCKASGDAFSS


SWMNWVKQRPGQGLEWIGRIYLGDGDINYNGKFKGRATLTADKSSSTAYM


QLSSLTSVDSAVYFCARQLGLWYVMDYWGQGTSVTVSSAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


Anti-CD1d_2B5.3G10_K-V-hIgGK-C (SEQ ID NO: 26):


ATGAGTGTGCCCACTCAGGTCCTGGGGTTGCTGCTGCTGTGGCTTACAGG


TGCCAGATGTGACATCCAGATGGCTCAGTCTCCAGCCTCCCTATCTGCAT


CTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAGTGAGAATATTTAC


AGTTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAGCTCCT


GGTCTATAATGCAAAAACCTTAGCAGAAGGTGTGCCATCAAGGTTCAGTG


GCAGTGGATCAGGCACACAGTTTTCTCTGAAGATCAACAGCCTGCAGCCT


GAAGATTTTGGGAGTTATTACTGTCAACATCATTATGGTTTTCCGTGGAC


GTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-CD1d_2B5.3G10_K-V-hIgGK-C (SEQ ID NO: 91):


MSVPTQVLGLLLLWLTGARCDIQMAQSPASLSASVGETVTITCRASENIY


SYLAWYQQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFSLKINSLQP


EDFGSYYCQHHYGFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-CD1d_2H11.2G5_H-V-hIgG4H-C (SEQ ID NO: 27):


ATGAACTTCGGGCTCAGCTTGATTTTCCTTGTCCTCATTTTAAAAGGTGT


CCAGTGTGAGGTGCAGCTGGTGGAGTCTGGGGGAGACTTAGTGAAGCCTG


GAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGC


TATGGCATGTCTTGGGTTCGCCAGACTCCAGACAAGAGGCTGGAGTGGGT


CGCAGTCATTAGTAGTGGTGGAAGTTCCACCTTCTATCCAGACAGTGTGA


AGGGGCGATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTG


CAAATGAGCAGTCTGAAGTCTGAGGACACAGCCGTGTATTACTGTTCAAG


AGGAGGTTACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCT


CCGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCC


AGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTA


CTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCG


GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTC


AGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACAC


CTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTG


AGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAA


GGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCAT


GATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGG


AAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCAT


AATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT


GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGT


ACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACC


ATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCC


CCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGG


TCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG


CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG


CTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGG


AGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC


TACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-CD1d_2H11.2G5_H-V-hIgG4H-C (SEQ ID NO: 92):


MNFGLSLIFLVLILKGVQCEVQLVESGGDLVKPGGSLKLSCAASGFTFSS


YGMSWVRQTPDKRLEWVAVISSGGSSTFYPDSVKGRFTISRDNAKNTLYL


QMSSLKSEDTAVYYCSRGGYYFDYWGQGTTLTVSAAKTKGPSVFPLAPCS


RSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL


SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFE


GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVH


NAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT


ISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNG


QPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNH


YTQKSLSLSLGKAS


 


Anti-CD1d_2H11.2G5_K-V-hIgGK-C (SEQ ID NO: 28):


ATGAGGTTCCAGGTTCAGGTTCTGGGGCTCCTTCTGCTCTGGATATCAGG


TGCCCAGTGTGATGTCCAGATAACCCAGTCTCCATCTTATCTTGCTGCAT


CTCCTGGAGAAACCATTACTATTAATTGCAGGGCAAGCAAGACCATTAGC


AAATATTTAGCCTGGTATCAAGAGAAACCTGAGAAAACTGATAAGCTTCT


TATCTACTCTGGATCCACTTTGCAATCTGGAATTCCATCAAGGTTCAGTG


GCAGTGGATCTGGTACAGATTTCACTCTCACCATCAGTGGCCTGGAGCCT


GAAGATTTTGCAATGTATTACTGTCAACAGCATAATGAATACCCGTGGAC


GTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-CD1d_2H11.2G5_K-V-hIgGK-C (SEQ ID NO: 93):


MRFQVQVLGLLLLWISGAQCDVQITQSPSYLAASPGETITINCRASKTIS


KYLAWYQEKPEKTDKLLIYSGSTLQSGIPSRFSGSGSGTDFTLTISGLEP


EDFAMYYCQQHNEYPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-CD40_11B6.1C3_H-LV-hIgG4H-C (SEQ ID NO: 29):


ATGGGATGGAGCTGGATCTTTCTCTTTCTCCTGTCAGGAACTGCAGGTGT


CCTCTCTGAGGTCCAGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTG


GGGCTTCAGTGAAGATATCCTGCAAGGCTTCTGGTTACTCATTCACTGGC


TACTACATGCACTGGGTGAAGCAAAGCCATGTAAAGAGCCTTGAGTGGAT


TGGACGTATTAATCCTTACAATGGTGCTACTAGCTACAACCAGAATTTCA


AGGACAAGGCCAGCTTGACTGTAGATAAGTCCTCCAGCACAGCCTACATG


GAGCTCCACAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


AGAGGACTACGTCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCAG


CCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGC


ACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCC


CGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGC


ACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGC


GTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAA


CGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCA


AATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGA


CCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTC


CCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACC


CCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCC


AAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAG


CGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGT


GCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCC


AAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATC


CCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAG


GCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCG


GAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTT


CTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGA


ATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACA


CAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-CD40_11B6.1C3_H-LV-hIgG4H-C (SEQ ID NO: 94):


MGWSWIFLFLLSGTAGVLSEVQLQQSGPELVKPGASVKISCKASGYSFTG


YYMHWVKQSHVKSLEWIGRINPYNGATSYNQNFKDKASLTVDKSSSTAYM


ELHSLTSEDSAVYYCAREDYVYWGQGTTLTVSSAKTKGPSVFPLAPCSRS


TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS


VVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGG


PSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA


KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS


KAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP


ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYT


QKSLSLSLGKAS


 


Anti-CD40_11B6.1C3_K-LV-hIgGK-C (SEQ ID NO: 30):


ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGATGTTCTGGATTCCTGCTTC


CAGCAGTGATGTTGTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTC


TTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGCCTTGTACAC


AGTAATGGAAACACCTATTTACATTGGTACCTGCAGAAGCCAGGCCAGTC


TCCAAAGCTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAG


ACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCGCACTCAAGATCAGT


AGAGTGGAGGCTGAGGATCTGGGAGTTTATTTCTGCTCTCAAAGTACACA


TGTTCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-CD40_11B6.1C3_K-LV-hIgGK-C (SEQ ID NO: 95):


MKLPVRLLVLMFWIPASSSDVVMTQTPLSLPVSLGDQASISCRSSQSLVH


SNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFALKIS


RVEAEDLGVYFCSQSTHVPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-CD40_12B4.2C10_H-LV-hIgG4H-C (SEQ ID NO: 31):


ATGGAATGGAGTTGGATATTTCTCTTTCTTCTGTCAGGAACTGCAGGTGT


CCACTCTGAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTG


GGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACATTCACTGAC


TATGTTTTGCACTGGGTGAAACAGAAGCCTGGGCAGGGCCTTGAGTGGAT


TGGATATATTAATCCTTACAATGATGGTACTAAGTACAATGAGAAGTTCA


AAGGCAAGGCCACACTGACTTCAGACAAATCCTCCAGCACAGCCTACATG


GAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTACTGTGCAAG


GGGCTATCCGGCCTACTCTGGGTATGCTATGGACTACTGGGGTCAAGGAA


CCTCAGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCC


CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTG


CCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAG


GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA


GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGG


CACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGG


TGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCA


GCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACC


CAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGG


TGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT


GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAA


CAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGC


TGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCC


TCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACA


GGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCA


GCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG


TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT


GCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACA


AGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAG


GCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAA


AGCTAGCTGA


 


Anti-CD40_12B4.2C10_H-LV-hIgG4H-C (SEQ ID NO: 96):


MEWSWIFLFLLSGTAGVHSEVQLQQSGPELVKPGASVKMSCKASGYTFTD


YVLHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYM


ELSSLTSEDSAVYYCARGYPAYSGYAMDYWGQGTSVTVSSAKTKGPSVFP


LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS


GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP


APEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD


GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE


WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE


ALHNHYTQKSLSLSLGKAS


 


Anti-CD40_12B4.2C10_K-LV-v2-hIgGK-C


(SEQ ID NO: 32):


ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGG


TACCAGATGTGATATCCAGATGACACAGACTACATCCTCCCTGTCTGCCT


CTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGC


AATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCT


GATCTACTACACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTG


GCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAA


GAAGATATTGCCACTTACTTTTGCCATCATGGTAATACGCTTCCGTGGAC


GTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-CD40_12B4.2C10_K-LV-v2-hIgGK-C


(SEQ ID NO: 97):


MMSSAQFLGLLLLCFQGTRCDIQMTQTTSSLSASLGDRVTISCRASQDIS


NYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTISNLEQ


EDIATYFCHHGNTLPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-CD40_12E12.3F3_H-V-hIgG4H-C (SEQ ID NO: 33):


ATGAACTTGGGGCTCAGCTTGATTTTCCTTGTCCTTGTTTTAAAAGGTGT


CCAGTGTGAAGTGAAGCTGGTGGAGTCTGGGGGAGGCTTAGTGCAGCCTG


GAGGGTCCCTGAAACTCTCCTGTGCAACCTCTGGATTCACTTTCAGTGAC


TATTACATGTATTGGGTTCGCCAGACTCCAGAGAAGAGGCTGGAGTGGGT


CGCATACATTAATTCTGGTGGTGGTAGCACCTATTATCCAGACACTGTAA


AGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTG


CAAATGAGCCGGCTGAAGTCTGAGGACACAGCCATGTATTACTGTGCAAG


ACGGGGGTTACCGTTCCATGCTATGGACTATTGGGGTCAAGGAACCTCAG


TCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTGA


 


Anti-CD40_12E12.3F3_H-V-hIgG4H-C (SEQ ID NO: 98):


MNLGLSLIFLVLVLKGVQCEVKLVESGGGLVQPGGSLKLSCATSGFTFSD


YYMYWVRQTPEKRLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYL


QMSRLKSEDTAMYYCARRGLPFHAMDYWGQGTSVTVSSAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


Anti-CD40_12E12.3F3_K-LV-hIgGK-C (SEQ ID NO: 34):


ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGG


TACCAGATGTGATATCCAGATGACACAGACTACATCCTCCCTGTCTGCCT


CTCTAGGAGACAGAGTCACCATCAGTTGCAGTGCAAGTCAGGGCATTAGC


AATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCT


GATCTATTACACATCAATTTTACACTCAGGAGTCCCATCAAGGTTCAGTG


GCAGTGGGTCTGGGACAGATTATTCTCTCACCATCGGCAACCTGGAACCT


GAAGATATTGCCACTTACTATTGTCAGCAGTTTAATAAGCTTCCTCCGAC


GTTCGGTGGAGGCACCAAACTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-CD40_12E12.3F3_K-LV-hIgGK-C (SEQ ID NO: 99):


MMSSAQFLGLLLLCFQGTRCDIQMTQTTSSLSASLGDRVTISCSASQGIS


NYLNWYQQKPDGTVKLLIYYTSILHSGVPSRFSGSGSGTDYSLTIGNLEP


EDIATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_24A5.4A5_H-V-hIgG4H-C (SEQ ID NO: 35):


ATGGATTGGCTGTGGAACTTGCTATTCCTGATGGCAGCTGCCCAAAGTGC


CCAAGCACAGATCCAGTTGGTGCAGTCTGGACCTGAGCTGAAGAAGCCTG


GAGAGACAGTCAAGATCTCCTGCAAGGCTTCTGGGTATTCCTTCACAAAC


TATGGAATGAACTGGGTGAAACAGGCTCCAGGAAAGGGTTTAAAGTGGAT


GGGCTGGATAAACACCTACACTGGAGAGTCAACATATGCTGATGACTTCA


AGGGACGGTTTGCCTTCTCTTTGGAAACCTCTGCCAGCACTGCCTATTTG


CAGATCAGTAACCTCAAAAATGAGGACATGGCTACATATTTCTGTGCTAG


AGGGGACTTTAGGTACTACTATTTTGACTACTGGGGCCAAGGCACCACTC


TCACAGGCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTGAT


 


Anti-DCIR_24A5.4A5_H-V-hIgG4H-C (SEQ ID NO: 100):


MDWLWNLLFLMAAAQSAQAQIQLVQSGPELKKPGETVKISCKASGYSFTN


YGMNWVKQAPGKGLKWMGWINTYTGESTYADDFKGRFAFSLETSASTAYL


QISNLKNEDMATYFCARGDFRYYYFDYWGQGTTLTGSSAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


Anti-DCIR_24A5.4A5_K-V-hIgGK-C (SEQ ID NO: 36):


ATGAGTGTGCTCACTCAGGTCCTGGCGTTGCTGCTGCTGTGGCTTACAGG


TGCCAGATGTGACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCAT


CTGTGGGAGAAACTGTCACCATCACGTGTCGAGCAAGTGGGAATATTCAC


AATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAGCTCCT


GGTCTATAATGCAAAAACCTTGGCAGATGGTGTGCCATCAAGGTTCAGTG


GCAGTGGATCAGGAACACAATATTCTCTCAAGATCAACACCCTGCAGCCT


GAAGATTTTGGGAGTTATTACTGTCAACATTTTTGGGATTCTTGGACGTT


CGGTGGAGGCACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCATCTG


TCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCT


GTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTG


GAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAG


AGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTG


AGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCACCCA


TCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTT


AG


 


Anti-DCIR_24A5.4A5_K-V-hIgGK-C (SEQ ID NO: 101):


MSVLTQVLALLLLWLTGARCDIQMTQSPASLSASVGETVTITCRASGNIH


NYLAWYQQKQGKSPQLLVYNAKTLADGVPSRFSGSGSGTQYSLKINTLQP


EDFGSYYCQHFWDSWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTAS


VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL


SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_24E7.3H9_H-V-hIgG4H-C (SEQ ID NO: 37):


ATGGAATGGACCTGGGTCTTTCTCTTCCTCCTGTCAGTAACTGCAGGTGT


CCACTCCCAGGTTCAGCTGCAGCAGTCTGGAGCTGAGCTGATGAAGCCTG


GGGCCTCAGTGAAGATATCCTGCAAGGCTACTGGCTACACATTCAGTAGC


TACTGGATAGAGTGGGTAAAGCAGAGGCCTGGACATGGCCTTGAGTGGAT


TGGAGAGATTTTACCTGGAAGTGGTAGGACTAACGACAATGAGAAGTTCA


AGGGCAAGGCCACATTCACTGCAGATACATCCTCCAAGAAAGCCTACATG


CAACTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTATTGTGCAAG


AAGGGGTGGTTACTCCTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTG


TCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGC


TCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGA


CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCA


GCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCC


CTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTA


CACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAG


TTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTC


GAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCT


CATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCC


AGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG


CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCG


TGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGG


AGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAA


ACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCT


GCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC


TGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT


GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGA


CGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGC


AGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC


CACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-DCIR_24E7.3H9_H-V-hIgG4H-C (SEQ ID NO: 102):


MEWTWVFLFLLSVTAGVHSQVQLQQSGAELMKPGASVKISCKATGYTFSS


YWIEWVKQRPGHGLEWIGEILPGSGRTNDNEKFKGKATFTADTSSKKAYM


QLSSLTSEDSAVYYCARRGGYSFAYWGQGTLVTVSAAKTKGPSVFPLAPC


SRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS


LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEF


EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV


HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK


TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN


GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN


HYTQKSLSLSLGKAS


 


Anti-DCIR_24E7.3H9_K-V-hIgGK-C (SEQ ID NO: 38):


ATGACCATGTTCTCACTAGCTCTTCTCCTCAGTCTTCTTCTCCTCTGTGT


CTCTGATTCTAGGGCAGAAACAACTGTGACCCAGTCTATGACCATGTTCT


CACTAGCTCTTCTCCTCAGTCTTCTTCTCCTCTGTGTCTCTGATTCTAGG


GCAGAAACAACTGTGACCCAGTCTCCAGCATCCCTGTCCATGGCTATAGG


GGAAAAAGTCACCATCAGATGCGTAACCAGCACTGATATTGATGATGATG


TGAACTGGTACCAGCAGAAGCCAGGGGAACCTCCTAAACTCCTTATTTCA


GAAGGCAATACTCTTCGTCCTGGAGTCCCATCCCGATTCTCCAGCAGTGG


CTATGGTACAGATTTTGTTTTTACAATTGAGAACATGCTCTCAGAAGATG


TTGCAGATTACTACTGTTTGCAAAGTGGTAACTTGCCGTACACGTTCGGA


GGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCATCTGTCTT


CATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTG


TGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAG


GTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCA


GGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCA


AAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCACCCATCAG


GGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAGCC


AGCATCCCTGTCCATGGCTATAGGGGAAAAAGTCACCATCAGATGCGTAA


CCAGCACTGATATTGATGATGATGTGAACTGGTACCAGCAGAAGCCAGGG


GAACCTCCTAAACTCCTTATTTCAGAAGGCAATACTCTTCGTCCTGGAGT


CCCATCCCGATTCTCCAGCAGTGGCTATGGTACAGATTTTGTTTTTACAA


TTGAGAACATGCTCTCAGAAGATGTTGCAGATTACTACTGTTTGCAAAGT


GGTAACTTGCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACG


AACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGT


TGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCC


AGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA


CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCC


TCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTC


TATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAG


CTTCAACAGGGGAGAGTGTTAG


 


Anti-DCIR_24E7.3H9_K-V-hIgGK-C (SEQ ID NO: 103):


MTMFSLALLLSLLLLCVSDSRAETTVTQSPASLSMAIGEKVTIRCVTSTD


IDDDVNWYQQKPGEPPKLLISEGNTLRPGVPSRFSSSGYGTDFVFTIENM


LSEDVADYYCLQSGNLPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSG


TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST


LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_29E9.2E2_H-VhIgG4H-C (SEQ ID NO: 39):


ATGGCTTGGGTGTGGACCTTGCTATTCCTGATGGCAGCTGCCCAAAGTGC


CCAAGCACAGATCCAGTTGGTGCAGTCTGGACCTGAGCTGAAGAAGCCTG


GAGAGACAGTCAAGATCTCCTGCAAGGCTTCTGGGTATACCTTCACAAAC


TATGGAATGAACTGGGTGAAGCAGGCTCCAGGAAAGGGTTTAAAGTGGGT


GGGCTGGATAAACACCTTCACTGGAGAGCCAACATATGTTGATGACTTCA


AGGGACGGTTTGCCTTCTCTTTGGAAACCTCTGCCAGCACTGCCTATTTG


CAGATCAACAACCTCAAAAATGAGGACACGGCTACATATTTCTGTGCAAG


AGGGAATTTTAGGTACTACTACTTTGACTACTGGGGCCAAGGCACCACTC


TCACAGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTGA


 


Anti-DCIR_29E9.2E2_H-VhIgG4H-C (SEQ ID NO: 104):


MAWVWTLLFLMAAAQSAQAQIQLVQSGPELKKPGETVKISCKASGYTFTN


YGMNWVKQAPGKGLKWVGWINTFTGEPTYVDDFKGRFAFSLETSASTAYL


QINNLKNEDTATYFCARGNFRYYYFDYWGQGTTLTVSSAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


Anti-DCIR_29E9.2E2_K-V-hIgGK-C (SEQ ID NO: 40):


ATGAGTGTGCTCACTCAGGTCCTGGCGTTGCTGCTGCTGTGGCTTACAGG


TGCCAGATGTGACATCCAGATGACTCAGTCCCCAGCCTCCCTATCTGCAT


CTGTGGGAGAAACTGTCACCATCACATGTCGAACAAGTGGGAATATTCGC


AATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAACTCCT


GGTCTATAATGCAAAAACCTTAGCAGATGGTGTGCCATCAAGGTTCGGTG


GCAGTGGATCAGGAACACAATATTCTCTCAAGATCAACAGCCTGCAGCCT


GAAGATTTTGGGAATTATTACTGTCAACATTTTTGGAGTAGTCCGTACAC


GTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-DCIR_29E9.2E2_K-V-hIgGK-C (SEQ ID NO: 105):


MSVLTQVLALLLLWLTGARCDIQMTQSPASLSASVGETVTITCRTSGNIR


NYLAWYQQKQGKSPQLLVYNAKTLADGVPSRFGGSGSGTQYSLKINSLQP


EDFGNYYCQHFWSSPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_29G10.3D9_H-V-hIgG4H-C (SEQ ID NO: 41):


ATGATGGGATGGAGCTATATCATCCTCTTTTTGGTAGCAACAGCTACAGA


TGTCCACTCCCAGGTCCAACTGCAGCAGCCTGGGGCTGAACTGGTGAAGC


CTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACC


AGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGAGAAGGCCTTGAGTG


GATTGGAGAGATTAATCCTAGCTACGGTCGTACTGACTACAATGAGAAGT


TCAAGAACAAGGCCACACTGACTGTAGCCAAATCCTCCAGCACAGCCTAC


ATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGC


AAGAGGAGATTACTACGGTAGTAGCTCGTTTGCTTACTGGGGCCAAGGGA


CTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCC


CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTG


CCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAG


GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA


GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGG


CACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGG


TGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCA


GCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACC


CAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGG


TGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT


GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAA


CAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGC


TGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCC


TCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACA


GGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCA


GCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG


TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT


GCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACA


AGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAG


GCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAA


AGCTAGCGGATGGAGCTATATCATCCTCTTTTTGGTAGCAACAGCTACAG


ATGTCCACTCCCAGGTCCAACTGCAGCAGCCTGGGGCTGAACTGGTGAAG


CCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCAC


CAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGAGAAGGCCTTGAGT


GGATTGGAGAGATTAATCCTAGCTACGGTCGTACTGACTACAATGGGAAG


TTCAAGAACAAGGCCACACTGACTGTAGCCAAATCCTCCAGCACAGCCTA


CATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTG


CAAGAGGAGATTACTACGGTAGTAGCTCGTTTGCTTACTGGGGCCAAGGG


ACTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCC


CCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCT


GCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCA


GGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTC


AGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGG


GCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAG


GTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCC


AGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAAC


CCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTG


GTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGA


TGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCA


ACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGG


CTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTC


CTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCAC


AGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTC


AGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGA


GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCG


TGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGAC


AAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGA


GGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTA


AAGCTAGCTGA


 


Anti-DCIR_29G10.3D9_H-V-hIgG4H-C (SEQ ID NO: 106):


MMGWSYIILFLVATATDVHSQVQLQQPGAELVKPGASVKLSCKASGYTFT


SYWMHWVKQRPGEGLEWIGEINPSYGRTDYNEKFKNKATLTVAKSSSTAY


MQLSSLTSEDSAVYYCARGDYYGSSSFAYWGQGTLVTVSAAKTKGPSVFP


LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS


GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP


APEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD


GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE


WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE


ALHNHYTQKSLSLSLGKAS


 


Anti-DCIR_29G10.3D9_K-Var1-V-hIgGK-C


(SEQ ID NO: 42):


ATGGATTTTCAAGTGCAGATTTTCAGCTTCCTGCTAATGAGTGCCTCAGT


CATAATGTCCAGGGGACAAATTGTTCTCACCCAGTCTCCAGCACTCATGT


CTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGCTCAAAT


ATAAGTTACATGTACTGGTACCAGCAGAAGCCAAGATCCTCCCCCAAACC


CTGGATTTATCTCACATCCAACCTGGCTTCTGGAGTCCCTGCTCGCTTCA


GTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAACCAGCAGCATGGAG


GCTGAAGATGCTGCCACTTATTGCTGCCAGCAGTGGAGTAGTAACCCACC


CACGTTCGGTGCTGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCAC


CATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT


GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT


ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTG


TCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTG


ACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGT


CACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG


AGTGTTAG


 


Anti-DCIR_29G10.3D9_K-Var1-V-hIgGK-C


(SEQ ID NO: 107):


MDFQVQIFSFLLMSASVIMSRGQIVLTQSPALMSASPGEKVTMTCSASSN


ISYMYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTTSSME


AEDAATYCCQQWSSNPPTFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGT


ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL


TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_29G10.3D9_K-Var2-V-hIgGK-C


(SEQ ID NO: 43):


ATGGATTTTCGAGTGCAGATTTTCAGCTTCCTGCTAATGAGTGCCTCAGT


CATAATGTCCAGGGGACAAATTGTTCTCACCCAGTCTCCAGCACTCATGT


CTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGCTCAAAT


ATAAGTTACATGTACTGGTACCAGCAGAAGCCAAGATCCTCCCCCAAACC


CTGGATTTATCTCACATCCAACCTGGCTTCTGGAGTCCCTGCTCGCTTCA


GTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAG


GCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCACC


CACGTTCGGTGCTGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCAC


CATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT


GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT


ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTG


TCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTG


ACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGT


CACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG


AGTGTTAG


 


Anti-DCIR_29G10.3D9_K-Var2-V-hIgGK-C


(SEQ ID NO: 108):


MDFRVQIFSFLLMSASVIMSRGQIVLTQSPALMSASPGEKVTMTCSASSN


ISYMYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSME


AEDAATYYCQQWSSNPPTFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGT


ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL


TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_2C9K-V-hIgGK-C (SEQ ID NO: 44):


ATGGAGACAGACACACTCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGG


TTCCACAGGTGACATTGTGCTGATCCAATCTCCAGCTTCTTTGGCTGTGT


CTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTGAT


AGTTATGTCAATAGTTTTATGCACTGGTACCAGCAGAAACCAGGACAGCC


ACCCAAACTCCTCATCTATCGTGTATCCAACCTAGAATCTGGGATCCCTG


CCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTAAT


CCTGTGGAGGCTGATGATGTTGCAACCTATTACTGTCAGCAAAGTAATGA


GGATCCATTCACGTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-DCIR_2C9K-V-hIgGK-C (SEQ ID NO: 109):


METDTLLLWVLLLWVPGSTGDIVLIQSPASLAVSLGQRATISCRASESVD


SYVNSFMHWYQQKPGQPPKLLIYRVSNLESGIPARFSGSGSRTDFTLTIN


PVEADDVATYYCQQSNEDPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.


 


Anti-DCIR_31A6.1F5_H-var2-V-hIgG4H-C


(SEQ ID NO: 45):


ATGGAATGTAACTGGATACTTCCTTTTATTCTGTCGGTAATTTCAGGGGT


CTACTCAGAGGTTCAGCTCCAGCAGTCTGGGACTGTGCTGGCAAGGCCTG


GGGCTTCCGTGAATATGTCCTGTAAGGCTGCTGGCTACAGCTTTACCAGT


TACTGGGTGTACTGGGTCAAACAGAGGCCTGGACAGGGTCTGGAATGGAT


TGGTGCTATTTACCCTAAAAATAGTAGAACTAGCTACAACCAGAAGTTCC


AGGACAAGGCCACACTGACTGCAGTCACATCCGCCAGCACTGCCTACATG


GAGCTCAGCAGCCTGACAAATGAGGACTCTGCGGTCTATTACTGTACAAG


ACCTCACTATGATTCGTTTGGTTACTGGGGCCAAGGGACTCTGGTCACTG


TCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGC


TCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGA


CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCA


GCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCC


CTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTA


CACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAG


TTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTC


GAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCT


CATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCC


AGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG


CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCG


TGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGG


AGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAA


ACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCT


GCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC


TGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT


GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGA


CGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGC


AGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC


CACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-DCIR_31A6.1F5_H-var2-V-hIgG4H-C


(SEQ ID NO: 110):


MECNWILPFILSVISGVYSEVQLQQSGTVLARPGASVNMSCKAAGYSFTS


YWVYWVKQRPGQGLEWIGAIYPKNSRTSYNQKFQDKATLTAVTSASTAYM


ELSSLTNEDSAVYYCTRPHYDSFGYWGQGTLVTVSAAKTKGPSVFPLAPC


SRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS


LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEF


EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV


HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK


TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN


GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN


HYTQKSLSLSLGKAS


 


Anti-DCIR_31A6.1F5_K-var2-V-hIgGK-C


(SEQ ID NO: 46):


ATGGAGACAGACACACTCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGG


TTCCACAGGTGACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGT


CTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTAGAT


AGTTATGGCATTAGTTTTATGCACTGGTACCAGCAGAAACCAGGACAGCC


ACCCAAACTCCTCATCTATCGTGCATCCAACCAAGAATCTGGGATCCCTG


CCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTAAT


CCTGTGGAGGCTGATGATGTTGCAACCTATTACTGTCAGCAAAGTAATGA


GGATCCGCTCACGTTCGGTGCTGGGACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-DCIR_31A6.1F5_K-var2-V-hIgGK-C


(SEQ ID NO: 111):


METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISCRASESVD


SYGISFMHWYQQKPGQPPKLLIYRASNQESGIPARFSGSGSRTDFTLTIN


PVEADDVATYYCQQSNEDPLTFGAGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_3C2.2D9_H-LV-hIgG4H-C (SEQ ID NO: 47):


ATGAACAGGCTTACTTCCTCATTGCTGCTGCTGATTGTCCCTGCATATGT


CCTGTCCCAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCT


CCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAGCACT


TCTGGTATGGGTGTGAGCTGGATTCGTCAGCCTTCAGGAAAGGGTCTGGA


GTGGCTGGCACACATTTACTGGGATGATGACAAGCGCTATAATCCATCCC


TGAAGAGCCGGCTCACAATCTTTAAGGATCCCTCCAGCAACCAGGTATTC


CTCAGGATCACCAGTGTGGACACTGCAGATACTGCCACATACTACTGTGC


TCGAAACTCCCATTACTACGGTAGTACTTACGGGGGATACTTCGATGTCT


GGGGCGCAGGGACCACGGTCACCGTCTCCTCAGCCAAAACAAAGGGCCCA


TCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGC


CGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT


CGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTC


CTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTC


CAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCA


GCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGC


CCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTT


CCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCA


CGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAAC


TGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA


GGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGC


ACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAA


GGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC


CCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCA


AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGAC


ATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGAC


CACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGC


TAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCC


GTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCT


GTCTCTGGGTAAAGCTAGCTGA


 


Anti-DCIR_3C2.2D9_H-LV-hIgG4H-C (SEQ ID NO: 112):


NRLTSSLLLLIVPAYVLSQQVTLKESGPGILQPSQTLSLTCSFSGFSLST


SGMGVSWIRQPSGKGLEWLAHIYWDDDKRYNPSLKSRLTIFKDPSSNQVF


LRITSVDTADTATYYCARNSHYYGSTYGGYFDVWGAGTTVTVSSAKTKGP


SVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV


LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC


PPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFN


WYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK


GLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSD


IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS


VMHEALHNHYTQKSLSLSLGKAS.


 


Anti-DCIR_3C2.2D9_K-LV-hIgGK-C (SEQ ID NO: 48):


ATGGAGACAGACACACTCCTGCTATGGGTGCTGCTGCTCGGGGTTCCAGG


TTCCACAGGTAACATTGTGCTGACCCAGTCTCCAACTTCTTTCACTGTGT


CTCTTGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTCAT


AGTTATGGCAATAGTTTTATGCACTGGTACCAGCAGAAACCAGGGCAGCC


ACCCAAACTCCTCATCTATCTTGCATCCAACGTAGAATCTGGGGTCCCTG


CCAGGTTCAGTGGTAGTGGGTCCAGGACAGACTTCACCCTCACCATTGAT


CCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAGTGA


GGATCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-DCIR_3C2.2D9_K-LV-hIgGK-C (SEQ ID NO: 113):


METDTLLLWVLLLGVPGSTGNIVLTQSPTSFTVSLGQRATISCRASESVH


SYGNSFMHWYQQKPGQPPKLLIYLASNVESGVPARFSGSGSRTDFTLTID


PVEADDAATYYCQQNSEDPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_6C8.1G9_H-V-hIgG4H-C (SEQ ID NO: 49):


ATGGAATGGACCTGGGTCTTTCTCTTCCTCCTGTCAGTAACTGCAGGTGT


CCACTCCCAGGTTCAGCTGCAGCAGTCTGGAACTGAGCTGATGAAGCCTG


GGGCCTCAGTGAAGATATCCTGCAAGGCTACTGGCTACACATTCAGTACC


TACTGGATAGAGTGGGTAAAGCAGAGGCCTGGACATGGCCTTGAGTGGAT


TGGAGAGATTTTACCTGGAAGTGGTAGGACTAACGACAATGAGAAGTTCA


AGGGCAAGGCCACAATCACTGCAGATACATCCTCCAAGAAAGCCTACATG


CAACTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTGCAAG


AAGGGGTGGTTACTCCTTTGCTTTCTGGGGCCAAGGGACTCTGGTCTCTG


TCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGC


TCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGA


CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCA


GCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCC


CTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTA


CACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAG


TTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTC


GAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCT


CATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCC


AGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG


CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCG


TGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGG


AGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAA


ACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCT


GCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC


TGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT


GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGA


CGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGC


AGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC


CACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-DCIR_6C8.1G9_H-V-hIgG4H-C (SEQ ID NO: 114):


MEWTWVFLFLLSVTAGVHSQVQLQQSGTELMKPGASVKISCKATGYTFST


YWIEWVKQRPGHGLEWIGEILPGSGRTNDNEKFKGKATITADTSSKKAYM


QLSSLTSEDSAVYYCARRGGYSFAFWGQGTLVSVSAAKTKGPSVFPLAPC


SRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS


LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEF


EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV


HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK


TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN


GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN


HYTQKSLSLSLGKAS


 


Anti-DCIR_6C8.1G9_K-V-hIgGK-C (SEQ ID NO: 50):


ATGACCATGTTCTCACTAGCTCTTCTCCTCAGTCTTCTTCTCCTCTGTGT


CTCTGATTCTAGGGCAGAAACAACTGTGACCCAGTCTCCAGCATCCCTGT


CCATGGCTATAGGAGAAAAAGTCACCATCAGATGCGTAACCAGCACTGAT


ATTGATGATGATGTGAACTGGTACCAGCAGAAGCCAGGGGAACCTCCTAA


GCTCCTTATTTCAGAAGGCAATACTCTTCGTGCTGGAGTCCCATCCCGAT


TCTCCAGCAGTGGCTATGGTACAGATTTTGTTTTTACAATTGAGAACATG


CTCTCAGAAGATGTTGCAGATTACTACTGTTTGCAAAGTGGTAACTTGCC


GTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTG


CACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGA


ACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAA


AGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGA


GTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACC


CTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGA


AGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGG


GAGAGTGTTAG


 


Anti-DCIR_6C8.1G9_K-V-hIgGK-C (SEQ ID NO: 115):


MTMFSLALLLSLLLLCVSDSRAETTVTQSPASLSMAIGEKVTIRCVTSTD


IDDDVNWYQQKPGEPPKLLISEGNTLRAGVPSRFSSSGYGTDFVFTIENM


LSEDVADYYCLQSGNLPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSG


TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST


LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR_9E8.1E3_H-V-hIgG4H-C (SEQ ID NO: 51):


ATGAACAGGCTTACTTCCTCATTGCTGCTGCTGATTGTCCCTGCATATGT


CCTGTCCCAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCT


CCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAGCACT


TCTGGTATGGGTCTGAGCTGGATTCGTCAGCCTTCAGGAAAGGGTCTGGA


GTGGCTGGCACACATTTACTGGGATGATGACAAGCGCTATAACCCATCCC


TGAAGAGCCGGCTCACAATCTCCAAGGATACCTCCAGCAACCAGGTTTTC


CTCAAGATCACCATTGTGGACACTGCAGATGCTGCCACATACTACTGTGC


TCGAAGCTCCCATTACTACGGTTATGGCTACGGGGGATACTTCGATGTCT


GGGGCGCAGGGACCACGGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCA


TCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGC


CGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT


CGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTC


CTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTC


CAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCA


GCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGC


CCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTT


CCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCA


CGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAAC


TGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA


GGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGC


ACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAA


GGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC


CCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCA


AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGAC


ATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGAC


CACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGC


TAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCC


GTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCT


GTCTCTGGGTAAAGCTAGCTGA


 


Anti-DCIR_9E8.1E3_H-V-hIgG4H-C (SEQ ID NO: 116):


MNRLTSSLLLLIVPAYVLSQVTLKESGPGILQPSQTLSLTCSFSGFSLST


SGMGLSWIRQPSGKGLEWLAHIYWDDDKRYNPSLKSRLTISKDTSSNQVF


LKITIVDTADAATYYCARSSHYYGYGYGGYFDVWGAGTTVTVSSAKTKGP


SVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV


LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC


PPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFN


WYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK


GLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSD


IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS


VMHEALHNHYTQKSLSLSLGKAS


 


Anti-DCIR_9E8.1E3_K-LV-hIgGK-C (SEQ ID NO: 52):


ATGGAGACAGACACACTCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGG


TTCCACAGGTAACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGT


CTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTATTCAT


AGTTATGGCAATAGTTTTCTGCACTGGTACCAGCAGAAACCAGGACAGCC


ACCCAAACTCCTCATCTATCTTGCATCCAACCTAGAATCTGGGGTCCCTG


CCAGGTTCAGCGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGAT


CCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAATGA


GGATCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAGGCGGCCGCACTAGCGCGGGCCGCATTCGAAGAG


CTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTGGC


CGCGACTCTAGATCATAATCAGC


 


Anti-DCIR_9E8.1E3_K-LV-hIgGK-C (SEQ ID NO: 117):


METDTLLLWVLLLWVPGSTGNIVLTQSPASLAVSLGQRATISCRASESIH


SYGNSFLHWYQQKPGQPPKLLIYLASNLESGVPARFSGSGSRTDFTLTID


PVEADDAATYYCQQNNEDPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-DCIR2C9H-LV-hIgG4H-V-hIgG4H-C (SEQ ID NO: 53):


ATGAAATGCAGCTGGGTCATCTTCTTCCTGATGGCAGTGGTTACAGGGGT


CAATTCAGAGGTTCAGCTGCAGCAGTCTGGGGCTGAGCTTGTGAGGCCAG


GGGCCTTAGTCAAGTTGTCCTGCAAAGCTTCTGGCTTCAACATTAATGAC


TACTATATCCACTGGGTGAAGCAGCGGCCTGAACAGGGCCTGGAGCGGAT


TGGATGGATTGATCCTGACAATGGTAATACTATATATGACCCGAAGTTCC


AGGGCAAGGCCAGTATAACAGCAGACACATCCCCCAACACAGCCTACCTG


CAGCTCAGCAGCCTGACATCTGAGGACACTGCCGTCTATTACTGTGCTAG


AACCCGATCTCCTATGGTTACGACGGGGTTTGTTTACTGGGGCCAAGGGA


CTGTGGTCACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCCCC


CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTG


CCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAG


GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA


GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGG


CACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGG


TGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCA


GCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACC


CAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGG


TGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT


GGCGTGGAGGTGCATAATGCCAAGACRAAGCCGCGGGAGGAGCAGTTCAA


CAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGC


TGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCC


TCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACA


GGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCA


GCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG


TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT


GCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACA


AGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAG


GCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAA


ATGA


 


Anti-DCIR2C9H-LV-hIgG4H-V-hIgG4H-C


(SEQ ID NO: 118):


MKCSWVIFFLMAVVTGVNSEVQLQQSGAELVRPGALVKLSCKASGFNIND


YYIHWVKQRPEQGLERIGWIDPDNGNTIYDPKFQGKASITADTSPNTAYL


QLSSLTSEDTAVYYCARTRSPMVTTGFVYWGQGTVVTVSAAKTKGPSVFP


LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS


GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP


APEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD


GVEVHNAKXKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE


WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE


ALHNHYTQKSLSLSLGK


 


Anti-DC-SIGNL16E3H (SEQ ID NO: 54):


ATGGAAAGGCACTGGATCTTTCTCTTCCTGTTTTCAGTAACTGCAGGTGT


CCACTCCCAGGTCCAGCTTCAGCAGTCTGGGGCTGAGCTGGCAAAACCTG


GGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTACTACC


TACTGGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGAT


TGGATACATTAATCCTATCACTGGTTATACTGAGTACAATCAGAAGTTCA


AGGACAAGGCCACCTTGACTGCAGACAAATCCTCCAGCACAGCCTACATG


CAACTGAGCAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


AGAGGGTTTAAGTGCTATGGACTATTGGGGTCAGGGAACCTCAGTCACCG


TCACCTCAGCCAAAACAACAGCCCCATCGGTCTATCCACTGGCCCCTGTG


TGTGGAGATACAACTGGCTCCTCGGTAACTCTAGGATGCCTGGTCAAGGG


TTATTTCCCTGAGCCAGTGACCTTGACCTGGAACTCTGGATCCCTGTCCA


GTGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGACCTCTACACCCTC


AGCAGCTCAGTGACTGTAACCTCGAGCACCTGGCCCAGCCAGACCGTCAC


CTGCAGCGTTGCTCACCCAGCCAGCAGCACCACGGTGGACAAAAAACTTG


AGCCCAGCGGGCCCATTTCAACAATCAACCCCTGTCCTCCATGCAAGGAG


TGTCACAAATGCCCAGCTCCTAACCTCGAGGGTGGACCATCCGTCTTCAT


CTTCCCTCCAAATATCAAGGATGTACTCATGATCTCCCTGACACCCAAGG


TCACGTGTGTGGTGGTGGATGTGAGCGAGGATGACCCAGACGTCCAGATC


AGCTGGTTTGTGAACAACGTGGAAGTACACACAGCTCAGACACAAACCCA


TAGAGAGGATTACAACAGTACTATCCGGGTGGTCAGCACCCTCCCCATCC


AGCACCAGGACTGGATGAGTGGCAAGGAGTTCAAATGCAAGGTCAACAAC


AAAGACCTCCCATCACCCATCGAGAGAACCATCTCAAAAATTAAAGGGCT


AGTCAGAGCTCCACAAGTATACATCTTGCCGCCACCAGCAGAGCAGTTGT


CCAGGAAAGATGTCAGTCTCACTTGCCTGGTCGTGGGCTTCAACCCTGGA


GACATCAGTGTGGAGTGGACCAGCAATGGGCATACAGAGGAGAACTACAA


GGACACCGCACCAGTCCTGGACTCTGACGGTTCTTACTTCATATATAGCA


AGCTCAATATGAAAACAAGCAAGTGGGAGAAAACAGATTCCTTCTCATGC


AACGTGAGACACGAGGGTCTGAAAAATTACTACCTGAAGAAGACCATCTC


CCGGTCTCCGGGTAAAGCTAGCTGA


 


Anti-DC-SIGNL16E3H (SEQ ID NO: 119):


MERHWIFLFLFSVTAGVHSQVQLQQSGAELAKPGASVKMSCKASGYTFTT


YWMHWVKQRPGQGLEWIGYINPITGYTEYNQKFKDKATLTADKSSSTAYM


QLSSLTSEDSAVYYCAREGLSAMDYWGQGTSVTVTSAKTTAPSVYPLAPV


CGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL


SSSVTVTSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPCKE


CHKCPAPNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVQI


SWFVNNVEVHTAQTQTHREDYNSTIRVVSTLPIQHQDWMSGKEFKCKVNN


KDLPSPIERTISKIKGLVRAPQVYILPPPAEQLSRKDVSLTCLVVGFNPG


DISVEWTSNGHTEENYKDTAPVLDSDGSYFIYSKLNMKTSKWEKTDSFSC


NVRHEGLKNYYLKKTISRSPGKAS


 


Anti-DC-SIGNL16E3K (SEQ ID NO: 55):


ATGGGCATCAAGATGGAGTCACGGATTCAGGCATTTGTATTCGTGTTTCT


CTGGTTGTCTGGTGTTGGCGGAGACATTGTGATGACCCAGTCTCACAAAT


TCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGT


CAGGATGTGACTTCTGCTGTAGCCTGGTATCAACAAAAACCAGGGCAATC


TCCTAAACTACTGATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTG


ATCGCTTCACAGGCAGTGGATCTGGGACAGATTATACTCTCACCATCAGC


AGTGGGCAGGCTGAAGACCTGGCACTTTATTACTGTCACCAATATTATAG


CGCTCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAGTCAAACGGGCTG


ATGCTGCACCAACTGTATCCATCTTCCCACCATCCAGTGAGCAGTTAACA


TCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTACCCCAAAGA


CATCAATGTCAAGTGGAAGATTGATGGCAGTGAACGACAAAATGGCGTCC


TGAACAGTTGGACTGATCAGGACAGCAAAGACAGCACCTACAGCATGAGC


AGCACCCTCACGTTGACCAAGGACGAGTATGAACGACATAACAGCTATAC


CTGTGAGGCCACTCACAAGACATCAACTTCACCCATCGTCAAGAGCTTCA


ATAGGAATGAGTGTTAG


 


Anti-DC-SIGNL16E3K (SEQ ID NO: 120):


MESRIQAFVFVFLWLSGVGGDIVMTQSHKFMSTSVGDRVSVTCKASQDVT


SAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDYTLTISSGQA


EDLALYYCHQYYSAPRTFGGGTKLEVKRADAAPTVSIFPPSSEQLTSGGA


SVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLT


LTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC


 


Anti-DC-SIGNL16E7H-LV-hIgG4H-C (SEQ ID NO: 56):


ATGGAAAGGCACTGGATCTTTCTCTTCCTGTTTTCAGTAACTGCAGGTGT


CCACTCCCAGGTCCAGCTTCAGCAGTCTGGGGCTGAGCTGGCAAAACCTG


GGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTACTACC


TACTGGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGAT


TGGATACATTAATCCTATCACTGGTTATACTGAGTACAATCAGAAGTTCA


AGGACAAGGCCACCTTGACTGCAGACAAATCCTCCAGCACAGCCTACATG


CAACTGAGCAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


AGAGGGTTTAAGTGCTATGGACTATTGGGGTCAGGGAACCTCAGTCACCG


TCACCTCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCGCCCTGC


TCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGA


CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCA


GCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCC


CTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTA


CACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAG


TTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTC


GAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCT


CATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCC


AGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG


CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCG


TGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGG


AGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAA


ACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCT


GCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC


TGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT


GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGA


CGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGC


AGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC


CACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-DC-SIGNL16E7H-LV-hIgG4H-C (SEQ ID NO: 121):


MERHWIFLFLFSVTAGVHSQVQLQQSGAELAKPGASVKMSCKASGYTFTT


YWMHWVKQRPGQGLEWIGYINPITGYTEYNQKFKDKATLTADKSSSTAYM


QLSSLTSEDSAVYYCAREGLSAMDYWGQGTSVTVTSAKTTGPSVFPLAPC


SRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS


LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEF


EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV


HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK


TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN


GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN


HYTQKSLSLSLGKAS.


 


Anti-DC-SIGNL16E7K-LV-hIgGK-C (SEQ ID NO: 57):


ATGGGCATCAAGATGGAGTCACAGATTCAGGCATTTGTATTCGTGTTTCT


CTGGTTGTCTGGTGTTGGCGGAGACATTGTGATGACCCAGTCTCACAAAT


TCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGT


CAGGATGTGACTTCTGCTGTAGCCTGGTATCAACAAAAACCAGGGCAATC


TCCTAAACTACTGATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTG


ATCGCTTCACAGGCAGTGGATCTGGGACAGATTATACTCTCACCATCAGC


AGTGGGCAGGCTGAAGACCTGGCACTTTATTACTGTCACCAATATTATAG


CGCTCCTCGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-DC-SIGNL16E7K-LV-hIgGK-C (SEQ ID NO: 122):


MESQIQAFVFVFLWLSGVGGDIVMTQSHKFMSTSVGDRVSVTCKASQDVT


SAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDYTLTISSGQA


EDLALYYCHQYYSAPRTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Dectin_1_11B6.4_H-V-hIgG4H-C (SEQ ID NO: 58):


ATGGCTGTCCTGGCACTACTCCTCTGCCTGGTGGCTTTCCCAACTTGTAC


CCTGTCCCAGGTGCAACTGAAGGAGTCAGGACCTGGCCTGGTGGCGCCCT


CACAGAGCCTGTCCATTACCTGCTCTGTCTCTGGGTTCTCATTAAGCAAC


TATGATATAAGCTGGATTCGCCAGCCACCAGGAAAGGGTCTGGAGTGGCT


TGGAGTAATGTGGACTGGTGGAGGCGCAAATTATAATTCAGCTTTCATGT


CCAGACTGAGCATCAACAAGGACAACTCCAAGAGCCAAGTTTTTTTAAAA


ATGAACAATCTGCAAACTGATGACACAGCCATTTATTACTGTGTCAGAGA


TGCGGTGAGGTACTGGAACTTCGATGTCTGGGGCGCAGGGACCACGGTCA


CCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCC


TGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAA


GGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGA


CCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTAC


TCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGAC


CTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGA


GAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAG


TTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACAC


TCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGA


GCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAG


GTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA


CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCA


AGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAG


AAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACAC


CCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCT


GCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC


AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTC


CGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGT


GGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC


AACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTG


A


 


Anti-Dectin_1_11B6.4_H-V-hIgG4H-C


(SEQ ID NO: 123):


MAVLALLLCLVAFPTCTLSQVQLKESGPGLVAPSQSLSITCSVSGFSLSN


YDISWIRQPPGKGLEWLGVMWTGGGANYNSAFMSRLSINKDNSKSQVFLK


MNNLQTDDTAIYYCVRDAVRYWNFDVWGAGTTVTVSSAKTKGPSVFPLAP


CSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY


SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPE


FEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE


VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE


KTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWES


NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH


NHYTQKSLSLSLGKAS


 


Anti-Dectin_1_11B6.4_K-LV-hIgGK-C (SEQ ID NO: 59):


ATGGATTTTCAAGCGCAGATTTTCAGCTTCCTGCTAATCAGTGCTTCAGT


CATAATGTCCAGAGGACAAATTGTTCTCTCCCAGTCACCAGCAATCCTGT


CTGCATCTCCAGGGGAGAAGGTCACAATGACTTGCAGGGCCAGCTCAAGT


GTAAGTTACATACACTGGTACCAGCAGAAGCCAGGATCCTCCCCCAAACC


CTGGATTTATGCCACATCCCACCTGGCTTCTGGAGTCCCTGCTCGCTTCA


GTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGAGTGGAG


GCTGAAGATACTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCATT


CACGTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAACTGTGGCTGCAC


CATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT


GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT


ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTG


TCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTG


ACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGT


CACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG


AGTGTTAG


 


Anti-Dectin_1_11B6.4_K-LV-hIgGK-C


(SEQ ID NO: 124):


MDFQAQIFSFLLISASVIMSRGQIVLSQSPAILSASPGEKVTMTCRASSS


VSYIHWYQQKPGSSPKPWIYATSHLASGVPARFSGSGSGTSYSLTISRVE


AEDTATYYCQQWSSNPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGT


ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL


TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Dectin_1_15E2.5_H-V-hIgG4H-C (SEQ ID NO: 60):


ATGGAAAGGCACTGGATCTTTCTACTCCTGTTGTCAGTAACTGCAGGTGT


CCACTCCCAGGTCCAGCTGCAGCAGTCTGGGGCTGAACTGGCAAGACCTG


GGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTACTACC


TACACTATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGAT


TGGATACATTAATCCTAGCAGTGGTTATACTAATTACAATCAGAAGTTCA


AGGACAAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGCCTCCATG


CAACTGAGCAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


AGAGAGGGCGGTATTAGTCCCCTATGCTATGGACTACTGGGGTCAAGGAA


CCTCAGTCACCGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCC


CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTG


CCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAG


GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA


GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGG


CACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGG


TGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCA


GCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACC


CAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGG


TGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT


GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAA


CAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGC


TGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCC


TCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACA


GGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCA


GCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG


TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT


GCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACA


AGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAG


GCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAA


AGCTAGCTGA


 


Anti-Dectin_1_15E2.5_H-V-hIgG4H-C


(SEQ ID NO: 125):


MERHWIFLLLLSVTAGVHSQVQLQQSGAELARPGASVKMSCKASGYTFTT


YTMHWVKQRPGQGLEWIGYINPSSGYTNYNQKFKDKATLTADKSSSTASM


QLSSLTSEDSAVYYCARERAVLVPYAMDYWGQGTSVTVSSAKTKGPSVFP


LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS


GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP


APEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD


GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE


WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE


ALHNHYTQKSLSLSLGKAS


 


Anti-Dectin_1_15E2.5_K-V-hIgGK-C (SEQ ID NO: 61):


ATGCATTTTCAAGTGCAGATTTTCAGCTTCCTGCTAATCAGTGCCTCAGT


CATAATGTCCAGAGGACAAATTGTTCTCACCCAGTCTCCAGCAGTCATGT


CTGCATCTCCAGGGGAGAAGGTCACCATAACCTGCACTGCCAGCTCAAGT


TTAAGTTACATGCACTGGTTCCAGCAGAAGCCAGGCACTTCTCCCAAACT


CTGGCTTTATAGCACATCCATCCTGGCTTCTGGAGTCCCTACTCGCTTCA


GTGGCAGTGGATCTGGGACCTCTTACTCTCTCACAATCAGCCGAATGGAG


GCTGAAGATGCTGCCACTTATTACTGCCAGCAAAGGAGTAGTTCCCCATT


CACGTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAACTGTGGCTGCAC


CATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT


GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT


ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTG


TCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTG


ACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGT


CACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG


AGTGTTAG


 


Anti-Dectin_1_15E2.5_K-V-hIgGK-C (SEQ ID NO: 126):


MHFQVQIFSFLLISASVIMSRGQIVLTQSPAVMSASPGEKVTITCTASSS


LSYMHWFQQKPGTSPKLWLYSTSILASGVPTRFSGSGSGTSYSLTISRME


AEDAATYYCQQRSSSPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGT


ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL


TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Dectin_1_2D8.2D4H-V-hIgG4H-C (SEQ ID NO: 62):


ATGGGATGGACCTGGATCTTTATTTTAATCCTGTCAGTTACTACAGGTGT


CCACTCTGAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGAGAAGCCTG


GCGCTTCAGTGAAGATATCCTGCAAGGCTTCTGGTTACTCCTTCACTGGC


TACAACATGAACTGGGTGAAACAGAGCAATGGAAAGAGCCTTGAGTGGAT


TGGAAATATTGATCCTTACTATGGTGATACTAACTACAACCAGAAGTTCA


AGGGCAAGGCCACATTGACTGTAGACAAATCCTCCAGCACAGCCTACATG


CACCTCAAGAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


ACCCTACGGTAGTGAGGCCTACTTTGCTTACTGGGGCCAAGGGACTCTGG


TCACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTGA


 


Anti-Dectin_1_2D8.2D4H-V-hIgG4H-C


(SEQ ID NO: 127):


MGWTWIFILILSVTTGVHSEVQLQQSGPELEKPGASVKISCKASGYSFTG


YNMNWVKQSNGKSLEWIGNIDPYYGDTNYNQKFKGKATLTVDKSSSTAYM


HLKSLTSEDSAVYYCARPYGSEAYFAYWGQGTLVTVSAAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


Anti-Dectin_1_2D8.2D4K-V-hIgGK-C (SEQ ID NO: 63):


ATGGTGTCCACTTCTCAGCTCCTTGGACTTTTGCTTTTCTGGACTTCAGC


CTCCAGATGTGACATTGTGATGACTCAGTCTCCAGCCACCCTGTCTGTGA


CTCCAGGAGATAGAGTCTCTCTTTCCTGCAGGGCCAGCCAGAGTATTAGC


GACTACTTACACTGGTATCAACAAAAATCACATGAGTCTCCAAGGCTTCT


CATCAAATATGCTGCCCAATCCATCTCTGGGATCCCCTCCAGGTTCAGTG


GCAGTGGATCAGGGTCAGATTTCACTCTCAGTATCAACGGTGTGGAACCT


GAAGATGTTGGAGTGTATTACTGTCAAAATGGTCACAGCTTTCCGTACAC


GTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-Dectin_1_2D8.2D4K-V-hIgGK-C (SEQ ID NO: 128):


DIVMTQSPATLSVTPGDRVSLSCRASQSISDYLHWYQQKSHESPRLLIKY


AAQSISGIPSRFSGSGSGSDFTLSINGVEPEDVGVYYCQNGHSFPYTFGG


GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV


DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG


LSSPVTKSFNRGEC


 


Anti-Langerin15B10H-LV-hIgG4H-C (SEQ ID NO: 64):


ATGGAATGGAGGATCTTTCTCTTCATCCTGTCAGGAACTGCAGGTGTCCA


CTCCCAGGTTCAGCTGCGGCAGTCTGGACCTGAGCTGGTGAAGCCTGGGG


CTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACATTTACTGACTAT


GTTATAAGTTGGGTGAAGCAGAGAACTGGACAGGGCCTTGAGTGGATTGG


AGATATTTATCCTGGAAGTGGTTATTCTTTCTACAATGAGAACTTCAAGG


GCAAGGCCACACTGACTGCAGACAAATCCTCCACCACAGCCTACATGCAG


CTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTGCAACCTA


CTATAACTACCCTTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCT


CTGCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCC


AGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTA


CTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCG


GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTC


AGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACAC


CTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTG


AGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAA


GGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCAT


GATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGG


AAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCAT


AATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT


GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGT


ACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACC


ATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCC


CCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGG


TCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG


CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGG


CTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGG


AGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC


TACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-Langerin15B10H-LV-hIgG4H-C (SEQ ID NO: 129):


QVQLRQSGPELVKPGASVKMSCKASGYTFTDYVISWVKQRTGQGLEWIGD


IYPGSGYSFYNENFKGKATLTADKSSTTAYMQLSSLTSEDSAVYFCATYY


NYPFAYWGQGTLVTVSAAKTTGPSVFPLAPCSRSTSESTAALGCLVKDYF


PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTC


NVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMI


SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV


SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPP


SQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS


FFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS.


 


Anti-Langerin15B10K-LV-hIgGK-C (SEQ ID NO: 65):


ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGATGTTCTGGATTCCTGCTTC


CAGCAGTGATGTTGTGATGACCCAAACTCCACTCTCCCTGCCTGTCCGTC


TTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGCCTTGTACAC


AGTAATGGAAACACCTATTTACATTGGTACCTGCAGAAGCCAGGCCAGTC


TCCAAAGCTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAG


ACAGGTTCAGTGGCAGTGGATCAGGGACAAATTTCACACTCAAGATCAGC


AGAGTGGAGGCTGAGGATCTGGGACTTTATTTCTGCTCTCAAAGTACACA


TGTTCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-Langerin15B10K-LV-hIgGK-C (SEQ ID NO: 130):


DVVMTQTPLSLPVRLGDQASISCRSSQSLVHSNGNTYLHWYLQKPGQSPK


LLIYKVSNRFSGVPDRFSGSGSGTNFTLKISRVEAEDLGLYFCSQSTHVP


YTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK


VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE


VTHQGLSSPVTKSFNRGEC


 


Anti-Langerin2G3H-LV-hIgG4H-C (SEQ ID NO: 66):


ATGACATTGAACATGCTGTTGGGGCTGAGGTGGGTTTTCTTTGTTGTTTT


TTATCAAGGTGTGCATTGTGAGGTGCAGCTTGTTGAGTCTGGTGGAGGAT


TGGTGCAGCCTAAAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTA


ACCTTCAATATCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGG


TTTGGAATGGGTTGCTCGCATAAGAAATAAAAGTAATAATTATGCAACAT


ATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA


CAAAGCTTGCTCTATCTGCAAATGAACAACTTGAAAACTGAGGACACAGC


CATGTATTACTGTGTGGGACGGGACTGGTTTGATTACTGGGGCCAAGGGA


CTCTGGTCACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCCCC


CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTG


CCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAG


GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCA


GGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGG


CACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGG


TGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCA


GCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACC


CAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGG


TGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT


GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAA


CAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGC


TGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCC


TCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACA


GGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCA


GCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG


TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT


GCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACA


AGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAG


GCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAA


AGCTAGCTGA


 


Anti-Langerin2G3H-LV-hIgG4H-C (SEQ ID NO: 131):


MTLNMLLGLRWVFFVVFYQGVHCEVQLVESGGGLVQPKGSLKLSCAASGL


TFNIYAMNWVRQAPGKGLEWVARIRNKSNNYATYYADSVKDRFTISRDDS


QSLLYLQMNNLKTEDTAMYYCVGRDWFDYWGQGTLVTVSAAKTKGPSVFP


LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS


GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP


APEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD


GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE


WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE


ALHNHYTQKSLSLSLGKAS


 


Anti-Langerin2G3L-LV-hIgGK-C (SEQ ID NO: 67):


ATGGCCTGGATTTCACTTATACTCTCTCTCCTGGCTCTCAGCTCAGGGGC


CATTTCCCAGGCTGTTGTGACTCAGGAATCTGCACTCACCACATCACCTG


GTGAAACAGTCACACTCACTTGTCGCTCAAGTACTGGGGCTGTTACAACT


AGTAACTATGCCAACTGGGTCCAAGAAAAACCAGATCATTTATTCACTGG


TCTAATAGGTGGTACCAACAACCGAGTTTCAGGTGTTCCTGCCAGATTCT


CAGGCTCCCTGATTGGAGACAAGGCTGCCCTCACCATCACAGGGGCACAG


ACTGAGGATGAGGCAATATATTTCTGTGCTCTATGGTACAGCAACCATTG


GGTGTTCGGTGGAGGAACCAAACTCGAGATCAAACGAACTGTGGCTGCAC


CATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT


GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT


ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTG


TCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTG


ACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGT


CACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG


AGTGTTAG


 


Anti-Langerin2G3L-LV-hIgGK-C (SEQ ID NO: 132):


MAWISLILSLLALSSGAISQAVVTQESALTTSPGETVTLTCRSSTGAVTT


SNYANWVQEKPDHLFTGLIGGTNNRVSGVPARFSGSLIGDKAALTITGAQ


TEDEAIYFCALWYSNHWVFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGT


ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL


TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Lox_1_10F9H-LV-hIgG4H-C (SEQ ID NO: 68):


ATGGAATGGACCTGGGTCTTTCTCTTCCTCCTGTCAGTAACTGCAGGTGT


CCACTCCCAGGTTCAGCTGCAGCAGTCTGGAGCTGAGCTGATGAAGCCTG


GGGCCTCAGTGAAGATATCCTGCAAGGCTACTGGCTACACATTCGGTAGC


TACTGGATAGAGTGGGTAAAGCAGAGGCCTGGACATGGCCTTGAGTGGAT


TGGAGAGATTTTACCTGGAAGTGGTAATACTAACTACAATGAGAACTTCA


AGGGCAAGGCCACATTCACTGCAGATACATCCTCCAACACAGCCTACATG


CAACTCACCAGTCTGACATCTGAGGACTCTGCCGTCTATTACTGTGCTAG


GGCGGGGATTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCA


AAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACC


TCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA


ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA


CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTG


GTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGT


AGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAAT


ATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCA


TCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCG


GACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCG


AGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAG


ACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGT


CCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCA


AGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAA


GCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCA


GGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCT


TCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAG


AACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTT


CCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATG


TCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAG


AAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-Lox_1_10F9H-LV-hIgG4H-C (SEQ ID NO: 133):


MEWTWVFLFLLSVTAGVHSQVQLQQSGAELMKPGASVKISCKATGYTFGS


YWIEWVKQRPGHGLEWIGEILPGSGNTNYNENFKGKATFTADTSSNTAYM


QLTSLTSEDSAVYYCARAGIYWGQGTLVTVSAAKTKGPSVFPLAPCSRST


SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV


VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGP


SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAK


TKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK


AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE


NNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ


KSLSLSLGKAS


 


Anti-Lox_1_10F9K-LV-hIgGK-C (SEQ ID NO: 69):


ATGGAGAAAGACACACTCCTGCTATGGGTCCTGCTTCTCTGGGTTCCAGG


TTCCACAGGTGACATTGTGCTGACCCAATCTCCAGCTTTTTTGGCTGTGT


CTCTAGGGCAGAGGGCCACCATCTCCTGCAGAGCCAGCGAAAGTGTTGAT


AATTATGGCATTAGTTTTATGAACTGGTTCCAACAGAAACCAGGACAGCC


ACCCAAACTCCTCATCTATGTTGCATCCAAGCAAGGATCCGGGGTCCCTG


CCAGGTTTAGTGGCAGTGGGTCTGGGACAGACTTCAGCCTCAACATCCAT


CCTATGGAGGAGGATGATACTGCAATGTATTTCTGTCAGCAAAGTAAGGA


GGTTCCTCGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-Lox_1_10F9K-LV-hIgGK-C (SEQ ID NO: 134):


MEKDTLLLWVLLLWVPGSTGDIVLTQSPAFLAVSLGQRATISCRASESVD


NYGISFMNWFQQKPGQPPKLLIYVASKQGSGVPARFSGSGSGTDFSLNIH


PMEEDDTAMYFCQQSKEVPRTFGGGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-LOX-111C8H-LV-hIgG4H-C (SEQ ID NO: 70):


ATGGAATGTAACTGGATACTTCCTTTTATTCTGTCGGTAACTTCAGGGGT


CTACTCAGAGGTTCAGCTCCAGCAGTCTGGGACTGTGCTGGCAAGGCCTG


GGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTACCAGC


TACTGGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGAT


TGGCGCTATTTATCCTGGAAATAGTGATACTACCTACAACCAGAAGTTCA


AGGGCAAGGCCAAACTGACTGCAGTCACATCCACCAGCACTGCCTACATG


GAGCTCAGCAGCCTGACAAATGAGGACTCTGCGGTCTATTACTGTACACC


TACTTACTACTTTGACTACTGGGGCCAAGGCACCTCTCTCACAGTCTCCT


CAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGG


AGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTT


CCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCG


TGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGC


AGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTG


CAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGT


CCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGG


GGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGAT


CTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAG


ACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAAT


GCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGT


CAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACA


AGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATC


TCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCC


ATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA


AAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAG


CCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTC


CTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGG


GGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTAC


ACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA


 


Anti-LOX-111C8H-LV-hIgG4H-C (SEQ ID NO: 135):


MECNWILPFILSVTSGVYSEVQLQQSGTVLARPGASVKMSCKASGYTFTS


YWMHWVKQRPGQGLEWIGAIYPGNSDTTYNQKFKGKAKLTAVTSTSTAYM


ELSSLTNEDSAVYYCTPTYYFDYWGQGTSLTVSSAKTKGPSVFPLAPCSR


STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS


SVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEG


GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHN


AKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI


SKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ


PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHY


TQKSLSLSLGKAS


 


Anti-LOX-111C8K-LV-hIgGK-C (SEQ ID NO: 71):


ATGAGTCCTGCCCAATTCCTGTTTCTGTTAGTGCTCTGGATTCGGGAAAC


CAACGGTGATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCA


TTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGAGCCTCTTAGAT


AGTGATGGAAAGACATATTTGAATTGGTTCTTACAGAGGCCAGGCCAGTC


TCCAAAGCGCCTAATCTATCTGGTGTCTAAACTGGACTCTGGAGTCCCTG


ACAGGTTCACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGC


AGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTACACA


TTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-LOX-111C8K-LV-hIgGK-C (SEQ ID NO: 136):


MSPAQFLFLLVLWIRETNGDVVMTQTPLTLSVTIGQPASISCKSSQSLLD


SDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKIS


RVEAEDLGVYYCWQGTHFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-LOX-115C4H-LV-hIgG4H-C (SEQ ID NO: 72):


ATGGGAGGGATCTGGATCTTTCTCTTCCTCCTGTCAGGAACTGCAGGTGC


CCACTCTGAGATCCAGCTGCAGCAGACTGGACCTGAGCTGGTGAAGCCTG


GGGCTTCAGTGAAGATATCCTGCAAGGCTTCTGGTTATCCATTCACTGAC


TACATCATGGTCTGGGTGAAGCAGAGCCATGGAAAGAGCCTTGAGTGGAT


TGGAAATATTAGTCCTTACTATGGTACTACTAACTACAATCTGAAGTTCA


AGGGCAAGGCCACATTGACTGTAGACAAATCTTCCAGCACAGCCTACATG


CAGCTCAACAGTCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAG


ATCCCCTAACTGGGACGGGGCCTGGTTTGCTCACTGGGGCCAAGGGGCTC


TGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTG


GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCT


GGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCG


CCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGA


CTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCAC


GAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGG


ACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCA


CCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAA


GGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGG


ACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGC


GTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAG


CACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA


ACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCC


ATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGT


GTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCC


TGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGG


GAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCT


GGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGA


GCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCT


CTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC


TAGCTGATTAATTAA


 


Anti-LOX-115C4H-LV-hIgG4H-C (SEQ ID NO: 137):


MGGIWIFLFLLSGTAGAHSEIQLQQTGPELVKPGASVKISCKASGYPFTD


YIMVWVKQSHGKSLEWIGNISPYYGTTNYNLKFKGKATLTVDKSSSTAYM


QLNSLTSEDSAVYYCARSPNWDGAWFAHWGQGALVTVSAAKTKGPSVFPL


APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG


LYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPA


PEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDG


VEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSS


IEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW


ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA


LHNHYTQKSLSLSLGKAS


 


Anti-LOX-115C4K-LV-hIgGK-C (SEQ ID NO: 73):


ATGGAGACAGACACAATCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGG


CTCCACTGGTGACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGT


CTCTAGGGCAGAGGGCCACCATCTCCTGCAAGGCCAGCCAAAGTGTTGAT


TATGATGGTGATAGTTATATGAACTGGTTCCAACAGAAACCAGGACAGCC


ACCCAAACTCCTCATCTATGCTGCATCCAATCTAGAATCTGGGATCCCAG


CCAGGTTTAGTGGCAGTGGGTCTGGGACAGACTTCACCCTCAACATCCAT


CCTGTGGAGGAGGAGGATGCTGCAACCTATTACTGTCAGCAAAGTAATGA


GGATCCATTCACGTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAACTG


TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA


TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGA


GGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCC


AGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC


AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC


CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA


ACAGGGGAGAGTGTTAG


 


Anti-LOX-115C4K-LV-hIgGK-C (SEQ ID NO: 138):


METDTILLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISCKASQSVD


YDGDSYMNWFQQKPGQPPKLLIYAASNLESGIPARFSGSGSGTDFTLNIH


PVEEEDAATYYCQQSNEDPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLK


SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS


STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Marco_10B7.3G4H-LV-hIgG4H-C (SEQ ID NO: 74):


ATGGCTGTCCTGGGGCTGCTTCTCTGCCTGGTGACGTTCCCAAGCTGTGT


CCTGTCCCAGGTGCAGCTGAAGGAGTCAGGACCTGGCCTGGTGGCACCCT


CACAGAGCCTGTCCATCACATGCACTGTCTCTGGGTTCTCATTATCCAGA


TATAGTGTATTTTGGGTTCGCCAGCCTCCAGGAAAGGGTCTGGAGTGGCT


GGGATTGATATGGGGTGGTGGAAGCACAGACTATAATTCAGCTCTCAAAT


CCAGACTGAGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAA


ATGAACAGTCTGCAAACTGATGACACAGCCATGTACTACTGTGCCAGAAT


CTACTTTGATTACGACGGGGCTATGGACTACTGGGGTCAAGGAACCTCAG


TCACCGTCTCCTCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTG


 


Anti-Marco_10B7.3G4H-LV-hIgG4H-C (SEQ ID NO: 139):


MAVLGLLLCLVTFPSCVLSQVQLKESGPGLVAPSQSLSITCTVSGFSLSR


YSVFWVRQPPGKGLEWLGLIWGGGSTDYNSALKSRLSISKDNSKSQVFLK


MNSLQTDDTAMYYCARIYFDYDGAMDYWGQGTSVTVSSAKTTGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


AntiMarco_10B7.3G4K_H-V-hIgGK-C (SEQ ID NO: 75):


ATGCATCGCACCAGCATGGGCATCAAGATGGAGTCACGGATTCAGGCATT


TGTATTCGTGTTTCTCTGGTTGTCTGGTGTTGGCGGAGACATTGTGATGA


CCCAGTCTCACAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTC


ACCTGCAAGGCCAGTCAGGATGTGACTTCTGCTGTAGCCTGGTATCAACA


AAAACCAGGGCAATCTCCTAAACTACTGATTTACTGGGCATCCACCCGGC


ACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTAT


ACTCTCACCATCAGCAGTGGGCAGGCTGAAGACCTGGCACTTTATTACTG


TCACCAATATTATAGCGCTCCTCGGACGTTCGGTGGAGGCACCAAGCTCG


AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCT


GATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAA


CTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCC


AATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGC


ACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAA


ACACAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCG


TCACAAAGAGCTTCAACAGGGGAGAGTGTTAG


 


AntiMarco_10B7.3G4K_H-V-hIgGK-C (SEQ ID NO: 140):


MHRTSMGIKMESRIQAFVFVFLWLSGVGGDIVMTQSHKFMSTSVGDRVSV


TCKASQDVTSAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDY


TLTISSGQAEDLALYYCHQYYSAPRTFGGGTKLEIKRTVAAPSVFIFPPS


DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDS


TYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Marco_11A8.3C9_H-V-hIgG4H-C (SEQ ID NO: 76):


ATGGAATGGAACTGGGTCGTTCTCTTCCTCCTGTCATTAACTGCAGGTGT


CTATGCCCAGGGTCAGATGCAGCAGTCTGGAGCTGAGCTGGTGAAGCCTG


GGGCTTCAGTGAAGCTGTCCTGCAAGACTTCTGGCTTCACCTTCAGCAGT


AACTATATAAGTTGGTTGAAGCAAAAGCCTGGACAGAGTCTTGAGTGGAT


TGCATGGATTTATGCTGGAACTGGTGGTATTACCTATAATCAGAAGTTCA


GAGGCAGGGCCCAACTGACTGTAGACACATCCTCCAGCACAGCCTACATG


CAGTTCAGCAGCCTGACAACTGATGACTCTGCCATCTATTACTGTGCAAG


ACACGTGAGGGGTTACCATCCTATGGACTACTGGGGTCAAGGAACCTCAG


TCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCG


CCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGT


CAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCC


TGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTC


TACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAA


GACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACA


AGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT


GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA


CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACG


TGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG


GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC


GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACG


GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATC


GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA


CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGA


CCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG


AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA


CTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCA


GGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG


CACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAG


CTGA


 


Anti-Marco_11A8.3C9_H-V-hIgG4H-C (SEQ ID NO: 141):


MEWNWVVLFLLSLTAGVYAQGQMQQSGAELVKPGASVKLSCKTSGFTFSS


NYISWLKQKPGQSLEWIAWIYAGTGGITYNQKFRGRAQLTVDTSSSTAYM


QFSSLTTDDSAIYYCARHVRGYHPMDYWGQGTSVTVSSAKTKGPSVFPLA


PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL


YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP


EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV


EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE


SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS


 


Anti-Marco_11A8.3C9_H-V-hIgGK-C (SEQ ID NO: 77):


ATGGAGTCACAGACTCAGGTCTTTGTATACATGTTGCTGTGGTTGTCTGG


TGTTGATGGAGACATTGTGATGACCCAGTCTCAAAAATTCATGTCCGCAT


CAGTAGGGGACAGGGTCAGCGTCACCTGCAGGGCCAGTCAGAATGTGGTT


ACTAATGTAGGCTGGTATCAACAGAAACCAGGGCAATCTCCTAAAGTACT


GATTTACTCGGCATCCTTCCGGTACAGTGGAGTCCCTGATCGCTTCACAG


GCAGTGGATCTGGGACAGATTTCACTCTCACCATCACCAATGTGCAGTCT


GAAGACTTGGCAGAGTATTTCTGTCAGCAATATAACAACTATCCGTACAC


GTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-Marco_11A8.3C9_H-V-hIgGK-C (SEQ ID NO: 142):


MESQTQVFVYMLLWLSGVDGDIVMTQSQKFMSASVGDRVSVTCRASQNVV


TNVGWYQQKPGQSPKVLIYSASFRYSGVPDRFTGSGSGTDFTLTITNVQS


EDLAEYFCQQYNNYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC


 


Anti-Marco_3H10.1F3_H-V-hIgG4H-C (SEQ ID NO: 78):


ATGGGATGGAGCTATATCATCCTCTTTTTGGTAGCAACAGCTACAGATGT


CCACTCCCAGGTCCAACTGCAGCAGCCTGGGGCTGAACTGGTGAAGCCTG


GGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGC


TACTGGATGCACTGGGTGAAGCAGAGGCCTGGAGAAGGCCTTGAGTGGAT


TGGAGAGATTAATCCTAGCTACGGTCGTACTGACTACAATGGGAAGTTCA


AGAACAAGGCCACACTGACTGTAGCCAAATCCTCCAGCACAGCCTACATG


CAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAG


AGGAGATTACTACGGTAGTAGCTCGTTTGCTTACTGGGGCCAAGGGACTC


TGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTG


GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCT


GGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCG


CCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGA


CTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCAC


GAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGG


ACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCA


CCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAA


GGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGG


ACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGC


GTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAG


CACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA


ACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCC


ATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGT


GTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCC


TGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGG


GAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCT


GGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGA


GCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCT


CTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC


TAGCTGA


 


Anti-Marco_3H10.1F3_H-V-hIgG4H-C (SEQ ID NO: 143):


MGWSYIILFLVATATDVHSQVQLQQPGAELVKPGASVKLSCKASGYTFTS


YWMHWVKQRPGEGLEWIGEINPSYGRTDYNGKFKNKATLTVAKSSSTAYM


QLSSLTSEDSAVYYCARGDYYGSSSFAYWGQGTLVTVSAAKTKGPSVFPL


APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG


LYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPA


PEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDG


VEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSS


IEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW


ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA


LHNHYTQKSLSLSLGKAS


 


Anti-Marco_3H10.1F3_K-V-hIgGK-C (SEQ ID NO: 79):


ATGGAGTCACAGACTCAGGTCTTTGTATACATGTTGCTGTGGTTGTCTGG


TGTTGATGGAGACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAT


CATTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGGT


ACTAATGTAGCCTGGTATCAACAGAAACCAGGGCACTCTCCTAAAGCACT


GATTTACTCGGCATCCTACCGGTACAGTGGAGTCCCTGATCGCTTCACAG


GCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCT


GAAGACTTGGCAGAGTTTTTCTGTCAGCAATATAACAACTATCCGTACAC


GTTCGGAGGGGGGACCACGCTCGAGATCAAACGAACTGTGGCTGCACCAT


CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC


TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA


GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA


CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG


CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCAC


CCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT


GTTAG


 


Anti-Marco_3H10.1F3_K-V-hIgGK-C (SEQ ID NO: 144):


MESQTQVFVYMLLWLSGVDGDIVMTQSQKFMSTSLGDRVSVTCKASQNVG


TNVAWYQQKPGHSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQS


EDLAEFFCQQYNNYPYTFGGGTTLEIKRTVAAPSVFIFPPSDEQLKSGTA


SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT


LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC






Humanized anti-CD40-HCV vaccine is: hAnti-CD40VK2-LV-hIgGK-C×hAnti-CD40VH3-LV-hIgG4H-C-Flex-v1-HelB-f1-ProtB-f2-NS5BPalm, wherein the portion of HelB are underlined, the portions of ProtB are bold and the portions of NS5BPalm are italicized. The linker sequence (in bold italics) is flanked by the transition sequence “AS” that bracket the linker sequences







[hAnti-CD40VK2-LV-hIgGK-C]







(SEQ ID NO: 158)







DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYY


TSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGG


GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK


VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV


THQGLSSPVTKSFNRGEC


 


hAnti-CD40VH3-LV-hIgG4H-C-Flex-v1-HelB-f1-ProtB-


f2-NS5BPalm







(SEQ ID NO: 159)







EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEW


VAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCA


RRGLPFHAMDYWGQGTLVTVSSAKTKGPSVFPLAPCSRSTSESTAAL


GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP


SSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGP


SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEV


HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI


EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW


ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV


MHEALHNHYTQKSLSLSLGKAS     ASVTVPHPNI


EEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVAL


GINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNTC


VTQTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGRGKPGIYRFVAP


GERAS   ASTPCTCGSSDLYLVTRHADVIPVRRRGDS


RGSLLSPRPISYLKGSSGGPLLCPAGHAVGIFRAAVCTRGVAKAVD


FIPVENLETTMRSPVFTDNSSPPAVPQSAS   ASVLDS


HYQDVLKEVKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPGQ


RVEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDIRTEEAIYQCCDL


DPQAFVAIKSLTERLYVGRCRASGVLTTSCGNTLTCYIKARAACRAA


GLQDCTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPPG


DPPQPEYDLELITAS






Humanized anti-DCIR-HCV 1st generation vaccine is: [hAnti-DCIRVK4-LV-hIgGK-C]×[hAnti-DCIRVH1-LV-hIgG4H-C-Flex-v1-HelB-f1-ProtB-f2-NS5BPalm] wherein the portion of HelB are underlined, the portions of ProtB are bold and the portions of NS5BPalm are italicized. The linker sequence (in bold italics) is flanked by the transition sequence “AS” that bracket the linker sequences







hAnti-DCIRVK4-LV-hIgGK-C







(SEQ ID NO: 160)







DIVMTQSPDSLAVSLGERATINCRASESIHSYGNSFLHWYQQKPGQPPKL


LIYLASNLESGVPSRFSGSGSRTDFTLTISSLQPEDFATYYCQQNNEDPW


TFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV


QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC


EVTHQGLSSPVTKSFNRGEC


 


hAnti-DCIRVH1-LV-hIgG4H-C-Flex-v1-HelB-f1-ProtB-


f2-NS5BPaLm







(SEQ ID NO: 161)







QVTLKESGPAIVKFTQTLTLTCSFSGFSLSTSGMGLSWIRQPSGKALEW


LAHIYWDDDKRYNPSLKSRLTISKDTSKNQVVLTMTIVDTVDAATYYCAR


SSHYYGYGYGGYFDVWGQGTTVTVSSAKTKGPSVFPLAPCSRSTSES


TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV


TVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEG


GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE


VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS


SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAV


EWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSC


SVMHEALHNHYTQKSLSLSLGKAS     ASVTV


PHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKL


VALGINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNT


CVTQTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGRGKPGIYRFVAP


GERAS   ASTPCTCGSSDLYLVTRHADVIPVRRRGDSR


GSLLSPRPISYLKGSSGGPLLCPAGHAVGIFRAAVCTRGVAKAVDFI


PVENLETTMRSPVFTDNSSPPAVPQSAS   SVLDSHYQ


DVLKEVKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPGQRVE


FLVQAWKSKKTPMGFSYDTRCFDSTVTESDIRTEEATYQCCDLDPQ


ARVAIKSLTERLYVGRCRASGVLTTSCGNTLTCYIKARAACRAAGLQD


CTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPPGDPPQ


PEYDLELITAS






Humanized anti-CD40-HCV vaccine is: hAnti-CD40VK2-LV-hIgGK-C-ViralHCVhelicasefgtB×hAnti-CD40VH3-LV-hIgG4H-C-Flex-v1-ProtB-f1-NS5BPalm, wherein the portion of ViralHCVhelicasefgtB are underlined, the portions of ProtB are bold and the portions of NS5BPalm are italicized. The linker sequence (in bold italics) is flanked by the transition sequence “AS” that bracket the linker sequences.







hAnti-CD40VK2-LV-hIgGK-C-ViralHCVhelicasefgtB







(SEQ ID NO: 162)







DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYY


TSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFG


GGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ


WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYA


CEVTHQGLSSPVTKSFNRGECASVTVPHPNIEEVALSTTGEIPFYGKA


IPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLDVSVIPTS


GVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTL


PQDAVSRTQRRGRTGRGKPGIYRFVAPGERAS


 


hAnti-CD40VH3-LV-hIgG4H-C-Flex-v1-ProtB-f1-


NS5BPalm







(SEQ ID NO: 163)







EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVA


YINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARR


GLPFHAMDYWGQGTLVTVSSAKTKGPSVFPLAPCSRSTSESTAALGC


LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS


LGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFL


FPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKT


KPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK


AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNG


QPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEAL


HNHYTQKSLSLSLGKAS     ASTPCTCGSSDLY


LVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGHAVGIF


RAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQSAS  


  ASVLDSHYQDVLKEVKAAASKVKANALYDVVSKLPLAVMG


SSYGFQYSPGQRVEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDIRTE


EAIYQCCDLDPQARVAIKSLTERLYVGRCRASGVLTTSCGNTLTCYIKAR


AACRAAGLQDCTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYS


APPGDPPQPEYDLELITAS






Humanized anti-DCIR-HCV 2nd generation vaccine is: [hAnti-DCIRVK4-LV-hIgGK-C-ViralHCVhelicasefgtB]×[hAnti-DCIRVH1-LV-hIgG4H-C-Flex-v1-ProtB-f1-NS5BPalm], wherein the portion of ViralHCVhelicasefgtB are underlined, the portions of ProtB are bold and the portions of NS5BPalm are italicized. The linker sequence (in bold italics) is flanked by the transition sequence “AS” that bracket the linker sequences.







hAnti-DCIRVK4-LV-hIgGK-C-ViralHCVhelicasefgtB







(SEQ ID NO: 164)







DIVMTQSPDSLAVSLGERATINCRASESIHSYGNSFLHWYQQKPGQPPKL


LIYLASNLESGVPSRFSGSGSRTDFTLTISSLQPEDFATYYCQQNNEDPW


TEGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV


QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC


EVTHQGLSSPVTKSENRGECASVTVPHPNIEEVALSTTGEIPFYGKAIPL


EVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLDVSVIPTSGVV


VVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTLPQDA


VSRTQRRGRTGRGKPGIYRFVAPGERAS


 


hAnti-DCIRVH1-LV-hIgG4H-C-Flex-v1-ProtB-f1-


NS5BPalm







(SEQ ID NO: 165)







QVTLKESGPAIVKPTQTLTLTCSFSGFSLSTSGMGLSWIRQPSGKALEWL


AHIYWDDDKRYNPSLKSRLTISKDTSKNQVVLTMTIVDTVDAATYYCARS


SHYYGYGYGGYFDVWGQGTTVTVSSAKTKGPSVFPLAPCSRSTSESTA


ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV


PSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGP


SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVH


NAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE


KTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW


ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV


MHEALHNHYTQKSLSLSLGKAS     ASTPCTC


GSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPA


GHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVETDNSSPPAVPQ


SAS     ASVLDSHYQDVLKEVKAAASKVKANA


LYDVVSKLPLAVMGSSYGFQYSPGQRVEFLVQAWKSKKTPMGESYD


TRCFDSTVTESDIRTEEATYQCCDLDPQARVAIKSLTERLYVGRCRAS


GVLTTSCGNTLTCYIKARAACRAAGLQDCTMLVCGDDLVVICESAGV


QEDAASLRAFTEAMTRYSAPPGDPPQPEYDLELITAS






Linkers can be a small as 2 amino acids, e.g., AS, but can also be longer, e.g., SSVSPTTSVHPTPTSVPPTPTKSSP (SEQ ID NO.: 166); PTSTPADSSTITPTATPTATPTIKG (SEQ ID NO.: 167); TVTPTATATPSAIVTTITPTATTKP (SEQ ID NO.: 168); TNGSITVAATAPTVTPTVNATPSAA (SEQ ID NO.: 169) or QTPTNTISVTPTNNSTPTNNSNPKPNP (SEQ ID NO:170).
It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method, kit, reagent, or composition of the invention, and vice versa. Furthermore, compositions of the invention can be used to achieve methods of the invention.
It will be understood that particular embodiments described herein are shown by way of illustration and not as limitations of the invention. The principal features of this invention can be employed in various embodiments without departing from the scope of the invention. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this invention and are covered by the claims.
All publications and patent applications mentioned in the specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and/or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and/or.” Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects.
As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.
The term “or combinations thereof” as used herein refers to all permutations and combinations of the listed items preceding the term. For example, “A, B, C, or combinations thereof” is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, expressly included are combinations that contain repeats of one or more item or term, such as BB, AAA, MB, BBC, AAABCCCC, CBBAAA, CABABB, and so forth. The skilled artisan will understand that typically there is no limit on the number of items or terms in any combination, unless otherwise apparent from the context.
All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.
REFERENCES
U.S. Patent Application Publication No. 2009/0238822: Chimeric HCV Antigens for Eliciting an Immune Response.
U.S. Patent Application Publication No. 2008/0241170: Vaccines Based on Targeting Antigen to DCIR Expressed on Antigen-Presenting Cells.
U.S. Patent Application Publication No. 2010/0239575: Anti-CD-40 Antibodies and Uses Thereof.

## Claims

1. A method for increasing effectiveness of Hepatitis C virus (HCV) antigen presentation by an antigen presenting cell (APC) comprising the steps of:
administering to a subject in need thereof a composition comprising an antibody conjugate that includes:
(i) a recombinant antibody having a light chain L and a heavy chain H, wherein said recombinant antibody binds to a CD40 receptor or a DCIR receptor;
(ii) two or three HCV antigens fused at a C-terminal position of said heavy chain H of said recombinant antibody;
(iii) one HCV antigen fused at a C-terminal position of said light chain L of said recombinant antibody;

wherein the antibody conjugate has a general structure given by:




wherein w, x, y, and z represent one or more HCV antigens or domains selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, or any combinations thereof.

2. The method of claim 1, wherein the antigen presenting cell comprises a dendritic cell (DC).

3. The method of claim 1, wherein the one or more native or engineered HCV antigenic peptide is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, E1b, and variants that have 95% identity with any one of the listed sequences.

4. The method of claim 1, wherein the DC-antibody is humanized.

5. The method of claim 1, wherein the composition further comprises one or more agents selected from the group consisting of an agonistic anti-CD40 antibody, an agonistic anti-CD40 antibody fragment, a CD40 ligand (CD40L) polypeptide, a CD40L polypeptide fragment, anti-4-1BB antibody, an anti-4-1BB antibody fragment, 4-1BB ligand polypeptide, a 4-1BB ligand polypeptide fragment, IFN-γ, TNF-α, type 1 cytokines, type 2 cytokines and combinations and modifications thereof.

6. The method of claim 1, wherein the HCV antigenic peptide is selected from the group consisting of protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof.

7. The method of claim 1, wherein the one or more HCV antigenic peptide is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and variants that have 95% identity with any one of the listed sequences.

8. The method of claim 1, wherein the HCV antigenic peptide is SEQ ID NO: 3 or an antigenic domain thereof.

9. The method of claim 1, wherein the antibody or antigen-binding fragment is selected from the group consisting of an antibody and antigen-binding fragment that binds to CD40.

10. The method of claim 1, wherein the antibody or antigen-binding fragment is selected from the group consisting of an antibody and antigen-binding fragment that binds to DCIR.

11. The method of claim 1, wherein the antibody conjugate comprises

12. The method of claim 1, wherein the antibody conjugate comprises

13. The method of claim 1, wherein the antibody conjugate has a general structure given by:



and wherein the antibody conjugate more effectively stimulates antigen presentation than an antibody conjugate having a general structure given by:

14. The method of claim 1, wherein the antibody conjugate has a general structure given by:



and wherein the antibody conjugate more effectively stimulates antigen presentation than an antibody conjugate having a general structure given by: