US20120301465A1

US20120301465A1 - Compositions and methods to immunize against hepatitis c virus

Info

Publication number: US20120301465A1
Application number: US13/430,206
Authority: US
Inventors: Helene Dutartre; Yves Levy; Jacques F. Banchereau; Gerard Zurawski
Original assignee: Institut National de la Sante et de la Recherche Medicale INSERM; Baylor Research Institute
Current assignee: Institut National de la Sante et de la Recherche Medicale INSERM; Baylor Research Institute
Priority date: 2011-03-25
Filing date: 2012-03-26
Publication date: 2012-11-29
Also published as: EP2688592A1; US20140199763A1; WO2012135132A1; TW201300418A; CA2831294A1; US9885017B2; AR085573A1; EP2688592A4

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.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application 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-linkerB-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, Fcγ 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 HelC, HelA, 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 IFN-γ-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 IFN-γ 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 IFN-γ 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 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: 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 GM-CSF, 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):
YQVRNSSGLYHVTNDCPNSSIVYEAADAILHTPGCVPCVREGNASRCWVAVTPTVATRDGKLPTTQ

LRRHIDLLVGSATLCSALYVGDLCGSVFLVGQLFTFSPRRHWTTQDCNCSIYPGHITGHRMAWDMM

MNWSPTTAVVAQLLRIPQAILDMIAGAHWGVLAGIAYFSMVGNWAKVLVVLLLFAGVDA

Envelop protein E2 (363 amino acids) (SEQ ID NO: 2):
ETHVTGGSAARTTAGLAGLFTPGAKQNIQLINTNGSWHINRTALNCNDSLNTGWVAGLFYYHKFNS

SGCPERLASCRPLTDFDQGWGPISYANGSGPDQRPYCWHYPPKPCGIVPAKSVCGPVYCFTPSPVVV

GTTDRSGAPTYNWGENDTDVFVLNNTRPPLGNWFGCTWMNSTGFTKVCGAPPCVIGGVGNNTLH

CPTDCFRKHPEATYSRCGSGPWITPRCLVDYPYRLWHYPCTINYTIFKIRMYVGGVEHRLEAACNW

TRGERCDLEDRDRSELSPLLLSTTQWQVLPCSFTTLPALSTGLIHLHQNIVDVQYLYGVGSSIASWAI

KWEYVVLLFLLLADARVCSCLWMMLLISQAEA

Non structural protein 3 NS3 (FL 631 amino acids) (SEQ ID NO: 3):
APITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQIVSTAAQTFLATCINGVCWTVYHGAGTRTIASPK

GPVIQMYTNVDQDLVGWPAPQGARSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYL

KGSSGGPLLCPAGHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQSFQVAHL

HAPTGSGKSTKVPAAYAAQGYKVLVLNPSVAATLGFGAYMSKAHGIDPNIRTGVRTITTGSPITYST

YGKFLADGGCSGGAYDIIICDECHSTDATSILGIGTVLDQAETAGARLVVLATATPPGSVTVPHPNIE

EVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLDVSVIPTSGV

VVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGRGKPGI

YRFVAPGERPSGMFDSSVLCECYDAGCAWYELTPAETTVRLRAYMNTPGLPVCQDHLEFWEGVFT

GLTHIDAHFLSQTKQSGENLPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPTPLLYRLG

AVQNEVTLTHPITKYIMTCMSADLEVVT

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

GPVIQMYTNVDQDLVGWPAPQGARSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYL

KGSSGGPLLCPAGHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQS

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

SPITYSTYGKFLADGGCSGGAYDIIICDECHSTDATSILGIGTVLDQAETAGARLVVLATATPPGSVTV

PHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVALGINAVAYYRGLDVSVI

PTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTLPQDAVSRTQRRGRTGR

GKPGIYRFVAPGERPSGMFDSSVLCECYDAGCAWYELTPAETTVRLRAYMNTPGLPVCQDHLEFW

EGVFTGLTHIDAHFLSQTKQSGENLPYLVAYQATVCARAQAPPPSWDQMWKCLIRLKPTLHGPTPL

LYRLGAVQNEVTLTHPITKYIMTCMSADLEVVT

Non structural NS5b (591 amino acids) (SEQ ID NO: 6):
SMSYSWTGALVTPCAAEEQKLPINALSNSLLRHHNLVYSTTSRSACQRQKKVTFDRLQVLDSHYQD

VLKEVKAAASKVKANLLSVEEACSLTPPHSAKSKFGYGAKDVRCHARKAVNHINSVWKDLLEDSV

TPIDTTIMAKNEVFCVQPEKGGRKPARLIVFPDLGVRVCEKMALYDVVSKLPLAVMGSSYGFQYSP

GQRVEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDIRTEEAIYQCCDLDPQARVAIKSLTERLYVGG

PLTNSRGENCGYRRCRASGVLTTSCGNTLTCYIKARAACRAAGLQDCTMLVCGDDLVVICESAGVQ

EDAASLRAFTEAMTRYSAPPGDPPQPEYDLELITSCSSNVSVAHDGAGKRVYYLTRDPTTPLARAA

WETARHTPVNSWLGNIIMFAPTLWARMILMTHFFSVLIARDQLEQALDCEIYGACYSIEPLDLPPIIQ

RLHGLSAFSLHSYSPGEINRVAACLRKLGVPPLRAWRHRARSVRARLLSRGGRAAICGKYLFNWAV

RTKLKLTPIAAAGQLDLSGWFTAGYSGGDIYHSVSHARPRWFWFCLLLLAAGVGIYLLPNR

The nucleotide sequences are presented herein below.

NS3Protease domain B (SEQ ID NO: 145)
ACTAGT ACTCCTTGTACCTGCGGCTCATCCGACCTGTACCTGGTCACCCGGCACGCAGACGTCA

TTCCTGTACGCCGACGCGGGGATAGTAGGGGGAGCCTGCTCTCTCCAAGACCCATATCCTACCT

CAAGGGCAGCAGCGGTGGACCACTGCTGTGTCCCGCTGGTCATGCTGTGGGAATATTTAGGGCC

GCAGTGTGTACCAGAGGCGTGGCCAAAGCTGTTGATTTTATTCCCGTCGAAAATCTTGAAACAA

CCATGAGAAGCCCAGTGTTCACAGACAACTCATCTCCCCCAGCAGTGCCGCAGAGT GCTAGC T

GAGAATTC GCGGCCGC

NS3Helicase domain B (SEQ ID NO: 146):
ACTAGT GTGACTGTGCCCCACCCCAATATCGAAGAGGTGGCCCTTAGTACTACCGGGGAAATTC

CTTTCTACGGGAAGGCCATCCCTCTCGAGGTTATTAAAGGAGGGCGACATCTGATTTTTTGCCA

CTCCAAGAAGAAGTGTGACGAGCTGGCCGCGAAACTGGTTGCCTTGGGCATCAACGCTGTCGC

ATACTATCGGGGACTGGATGTATCAGTGATACCCACCAGCGGAGTGGTAGTTGTCGTCGCTACA

GACGCATTGATGACCGGCTTTACAGGAGATTTCGACTCCGTCATCGACTGTAACACATGCGTGA

CTCAGACAGTGGATTTCAGCCTTGACCCGACGTTTACGATTGAGACCACCACTCTCCCTCAGGA

TGCTGTGTCTAGGACCCAAAGACGCGGTCGCACAGGCCGGGGCAAACCAGGCATCTATAGGTT

CGTGGCACCAGGGGAAAGA GCTAGC TGAgaattc GCGGCCGC

NS5bPalm (SEQ ID NO: 147):
ACTAGT GTGCTGGACTCTCACTACCAGGATGTCCTGAAGGAAGTAAAAGCAGCCGCTTCTAAA

GTCAAAGCGAACGCTCTGTACGATGTCGTTTCCAAACTGCCGCTGGCTGTCATGGGCTCTTCCTA

CGGCTTTCAGTATTCCCCGGGTCAGCGCGTTGAGTTCCTGGTCCAGGCGTGGAAATCCAAAAAG

ACTCCGATGGGTTTTTCCTATGACACTCGCTGCTTCGACAGCACCGTTACCGAAAGCGACATTC

GCACCGAGGAAGCAATCTACCAGTGCTGCGACCTGGACCCACAGGCCCGCGTGGCGATCAAAT

CTCTGACCGAACGCCTGTACGTTGGCCGCTGTCGCGCTTCCGGTGTTCTGACGACCTCCTGCGGT

AATACGCTGACCTGCTACATCAAAGCACGCGCTGCCTGTCGCGCAGCCGGTCTGCAGGACTGCA

CCATGCTGGTGTGTGGCGATGACCTGGTGGTGATCTGCGAAAGCGCTGGCGTGCAGGAAGACG

CAGCAAGCCTGCGCGCTTTCACCGAAGCTATGACTCGCTACTCTGCGCCGCCGGGTGACCCGCC

GCAGCCAGAATACGATCTGGAGCTGATCACC GCTAGC TAAGAATTC GCGGCCGC

SEQ ID NOS: 7-14 show the HCV antigen domains E1a, E2, ProtA, Prot B, Hel A, Hel B, HelC, 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

DQRPYCWHYPPKPCGIVPAKSVCGPVYCFTPSPVVVGTTDRSGAPTYNW

GENDTDVFVLNNTRPPLGNWFGCTWMNSTGFTKVCGAPPCVIGGVGNNT

LHCPTDCFRKHPEATYSRCGSGPWITPRCLVDYPYRLWHYPCTINYTI

FKIRMYVGGVEHRLEAACNWTRGERCDLEDRDRSELSPLLLSTTQWQVLP

CSFTTLPALSTGLIHLHQNIVDVQYLYGVGSSIASWAIKWEYVVLLFLL

AS

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)

TSAPITAYAQQTRGLLGCIITSLTGRDKNQVEGEVQIVSTAAQTFLATC

INGVCWTVYHGAGTRTIAS

Prot B

(SEQ ID NO: 10)

TSTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSG

GPLLCPAGHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNS

SPPAVPQSAS

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

VT AS

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 chromatograhy. 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 (Elutra™, 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 10⁶cells/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 mM each peptide) as indicated. Alternatively, TLR agonists (polylC, 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.

HCV

Viral

HLA

Patient ID

Genotype

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

Hispanic

after

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

Hispanic

responder

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

Hispanic

untreated

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

Hispanic

untreated

apheresis

HCV-VAC-026

3a

M

Non

White

positive

29

?

no

Hispanic

untreated

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 2×10⁶cells/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 PEcγ7-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 IFN-γ-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):
ATGGGCAGGCTTACTTCTTCATTCTTGCTACTGATTGTCCCTGCATATGTCCTGTCCCAGGTTACT

CTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACCTGTTCTTTCTC

TGGGTTTTCACTGAGCACTTCTGGTATGAGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGT

CTGGAGTGGCTGGCTCACATTTGGTGGAATGATGATAAGTACTATAATCCAGTCCTGAAAAGCC

GGCTCACAATCTCCAAGGAGACCTCCAACAACCAGGTATTCCTCAAGATCGCCAGTGTGGTCTC

TGCAGATACTGCCACATACTACTGTGCTCGATTCTATGGTAACTGTCTTGACTACTGGGGCCAA

GGCACCACTCTCACAGTCTCCTCGGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCT

GCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA

ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTC

CTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCA

CGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTG

AGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGT

CTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGC

GTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG

GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTC

AGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCC

AACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAG

CCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACC

TGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCG

GAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCA

GGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATG

AGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGATT

AATTAA

Anti_CLEC_6_9B9.2G12_Heavy (H)v-V-hIgG4H-C (SEQ ID NO: 80):
MGRLTSSFLLLIVPAYVLSQVTLKESGPGILQPSQTLSLTCSFSGFSLSTSGMSVGWIRQPSGKGLEWL

AHIWWNDDKYYNPVLKSRLTISKETSNNQVFLKIASVVSADTATYYCARFYGNCLDYWGQGTTLT

VSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL

SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMI

SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN

GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

CAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCA

GGGCAAGTCAGGACATTAGCAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTA

AACTCCTGATCTACTACACATCAATATTACAATTAGGAGTCCCATCAAGATTCAGTGGCAGTGG

GTCTGAAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTT

GCCAACAGGGTGATTCGCTTCCATTCACGTTCGGCTCGGGGACAAAGCTCGAGATCAAACGAAC

TGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCT

CTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAA

CGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTA

CAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTG

CGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

Anti_CLEC_6_9B9.2G12_Light (K)v-V-hIgGK-C (SEQ ID NO: 81):
MMSSAQFLGLLLLCFQGTRCDIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLL

IYYTSILQLGVPSRFSGSGSETDYSLTISNLEQEDIATYFCQQGDSLPFTFGSGTKLEIKRTVAAPSVFIF

PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA

DYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

TCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTCAGTCACTTTCACTCACCTGCACTGTCACTG

GCTACTCCATCACCAGTGGTTATAGCTGGCACTGGATCCGGCAGTTTCCAGGAAACAAACTGGA

ATGGATGGGCTACATACTCTTCAGTGGTAGCACTAACTACAACCCATCTCTGAAAAGTCGAATC

TCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGG

ACACAGCCACATATTTCTGTGCAAGATCTAACTATGGTTCCTTTGCTTCCTGGGGCCAAGGGACT

CTGGTCACTGTCTCTGCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCA

GGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGT

GACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAG

TCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGA

CCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCA

AATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCT

GTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTG

GTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG

CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTC

CTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAA

GGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAG

GTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG

GTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAAC

AACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAA

CCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCT

GCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGATTAATTAA

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

GYILFSGSTNYNPSLKSRISITRDTSKNQFFLQLNSVTTEDTATYFCARSNYGSFASWGQGTLVTVSA

AKTTGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV

VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRT

PEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYK

CKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE

NNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

ACAAATTGTTCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATG

ACCTGCAGTGCCAGCTCAAGTGTAAGTCACATGCACTGGTACCAGCAGAAGTCAGGCACTTCCC

CCAAAAGATGGATTTATGACACATCCAGACTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAG

TGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTAT

TACTGCCAGCAGTGGAGTAGTCACCCATGGTCGTTCGGTGGAGGCACCAAACTCGAGATCAAA

CGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAAC

TGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTG

GATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGC

ACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAT

GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAG

TGTTAG

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

YDTSRLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSHPWSFGGGTKLEIKRTVAAPSV

FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS

KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AGTTGGTGCAGTCTGGACCTGAGCTGAAGAAGCCTGGAGAGACAGTCAAGATCTCCTGCAAGG

CTTCTGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGTTCCAGGAAAAGGTTT

AAGGTGGATGGGCTGGATGGACACCTTCACTGGAGAGCCAACATATGCTGATGACTTCAAGGG

ACGGTTTGCCTTCTCTTTGGAAACCTCTGCCAGCACTGCCTATTTGCAGATCAACAGCCTCAAAA

ATGAGGACACGGCTACTTATTTCTGTGCAAGAGGGGGGATTTTACGACTCAACTACTTTGACTA

CTGGGGCCAAGGCACCACTCTCACAGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCC

CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACT

ACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTT

CCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC

AGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGAC

AAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGG

GACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGA

GGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGT

GGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA

CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGC

AAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAG

CCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTC

AGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT

GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC

TCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCG

TGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC

TAGCTGATTAATTAA

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

RWMGWMDTFTGEPTYADDFKGRFAFSLETSASTAYLQINSLKNEDTATYFCARGGILRLNYFDYW

GQGTTLTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL

QSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPP

KPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH

QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDI

AVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS

LSLGKAS

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

CCAGATGACACAATCTTCATCCTCCTTTTCTGTATCTCTAGGAGACAGAGTCACCATTACTTGCA

AGGCAAGTGAGGACATATATAATCGGTTAGGCTGGTATCAGCAGAAACCAGGAAATGCTCCTA

GGCTCTTAATATCTGGTGCAACCAGTTTGGAAACTGGGGTTCCTTCAAGATTCAGTGGCAGTGG

ATCTGGAAAGGATTACGCTCTCAGCATTACCAGTCTTCAGACTGAAGATCTTGCTACTTATTACT

GTCAACAGTGTTGGACTTCTCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAA

CTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC

TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATA

ACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCT

ACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCT

GCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTT

AG

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

GATSLETGVPSRFSGSGSGKDYALSITSLQTEDLATYYCQQCWTSPYTFGGGTKLEIKRTVAAPSVFI

FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA

DYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCT

TCTGGATACACCTTCACTGACTACTACATGAAGTGGGTGAAGCAGAGCCATGGAAAGAGCCTTG

AGTGGATTGGAGATATTAATCCTAACTATGGTGATACTTTCTACAACCAGAAGTTCGAGGGCAA

GGCCACATTGACTGTAGACAAATCCTCCAGGACAGCCTACATGCAGCTCAACAGCCTGACATCT

GAGGACTCTGCAGTCTATTATTGTGGAAGAGGGGACTATGGATACTTCGATGTCTGGGGCGCAG

GGACCACGGTCACCGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTG

CTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAA

CCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCC

TACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCAC

GAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGA

GTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTC

TTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCG

TGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGG

AGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCA

GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCA

ACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGC

CACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCT

GCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG

AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAG

GCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGA

GGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

WIGDINPNYGDTFYNQKFEGKATLTVDKSSRTAYMQLNSLTSEDSAVYYCGRGDYGYFDVWGAGT

TVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLLSLGKAS

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

TGTGATGACCCAGTCTCACAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCATCACCTGC

AAGGCCAGTCAGGATGTGGGTACTGCTGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCT

AAACTACTGATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCTTCACAGGCAGTG

GATCTGGGACAGATTTCACTCTCACCATTAACAATGTGCAGTCTGAAGACTTGGCAGATTATTT

CTGTCAGCAATATAGCAGCAATCCGTACATGTTCGGAGGGGGGACCAAGCTCGAGATCAAACG

AACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTG

CCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGA

TAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCAC

CTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC

CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG

TTAG

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

KLLIYWASTRHTGVPDRFTGSGSGTDFTLTINNVQSEDLADYFCQQYSSNPYMFGGGTKLEIKRTVA

APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST

LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAACAGTCTGGACCTGAGTTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTGCAAGACT

TCTGGATACACATTCACTGAATACACCATGCACTGGGTGAGGCAGAGCCATGGAAAGAGCCTT

GAGTGGATTGGAGGTATTAATCCTATCAATGGTGGTCCTACCTACAACCAGAAGTTCAAGGGCA

AGGCCACATTGACTGTTGACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCTGACATC

TGAGGACTCTGCAGTCTATTACTGTGCAAGATGGGACTATGGTAGTCGAGATGTTATGGACTAC

TGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCC

TGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTA

CTTCCCCGAACCGGTACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCAC

ACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTC

CAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGT

GGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAA

GGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCC

CTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGT

ACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAA

GTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGG

GCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC

AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCT

TCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA

GCTAGCTGA

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

GGINPINGGPTYNQKFKGKATLTVDKSSSTAYMELRSLTSEDSAVYYCARWDYGSRDVMDYWGQ

GTSVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVPVTVSWNSGALTSGVHTFPAVL

QSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPP

KPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH

QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDI

AVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS

LSLGKAS

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

TGTAATGACTCAATCTCCCAAATCCATGTCCATGTCAGTAGGGGAGAGGGTCACCTTGAGCTGC

AAGGCCAGTGAGAATGTGGGAACTTATGTATCCTGGTATCAACAGAGACCAGAACAGTCTCCA

AAACTGCTGATATACGGGGCATCCAACCGGTACACTGGGGTCCCCGATCGCTTCACAGGCAGTG

GATCTGCAACAGATTTCACTCTGACCATCAGCAGTGTGCAGGCTGAGGACCTTGCAGATTATCA

CTGTGGACAGACTTACAGCTATATATTCACGTTCGGCTCGGGGACAAAGCTCGAGATCAAACGA

ACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGC

CTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGAT

AACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACC

TACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCC

TGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

TAG

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

LLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQTYSYIFTFGSGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GGTGCAGCAGTCGGGACCTGAGTTGGTGAAGCCTGGGGCCTCAGTGAAGATTTCCTGCAAAGC

CTCTGGCGACGCATTCAGTAGTTCTTGGATGAACTGGGTGAAGCAGAGGCCTGGACAGGGTCTT

GAGTGGATTGGACGGATTTATCTTGGAGATGGAGATATTAATTACAATGGGAAGTTCAAGGGC

AGGGCCACACTGACTGCAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACCT

CTGTGGACTCTGCGGTCTATTTCTGCGCGAGGCAGCTCGGGCTATGGTATGTTATGGACTACTG

GGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTG

GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACT

TCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCC

GGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGC

TTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAG

AGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGAC

CATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTC

ACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT

GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGT

GTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAG

GTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCC

GAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCC

TGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC

AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTA

CAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGAT

GCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGC

TGA

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

WIGRIYLGDGDINYNGKFKGRATLTADKSSSTAYMQLSSLTSVDSAVYFCARQLGLWYVMDYWG

QGTSVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ

SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK

PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ

DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGKAS

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

TCCAGATGGCTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGT

CGAGCAAGTGAGAATATTTACAGTTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTC

AGCTCCTGGTCTATAATGCAAAAACCTTAGCAGAAGGTGTGCCATCAAGGTTCAGTGGCAGTGG

ATCAGGCACACAGTTTTCTCTGAAGATCAACAGCCTGCAGCCTGAAGATTTTGGGAGTTATTAC

TGTCAACATCATTATGGTTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAA

CTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC

TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATA

ACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCT

ACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCT

GCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTT

AG

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

LVYNAKTLAEGVPSRFSGSGSGTQFSLKINSLQPEDFGSYYCQHHYGFPWTFGGGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGGTGGAGTCTGGGGGAGACTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCC

TCTGGATTCACTTTCAGTAGCTATGGCATGTCTTGGGTTCGCCAGACTCCAGACAAGAGGCTGG

AGTGGGTCGCAGTCATTAGTAGTGGTGGAAGTTCCACCTTCTATCCAGACAGTGTGAAGGGGCG

ATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTGCAAATGAGCAGTCTGAAGTCT

GAGGACACAGCCGTGTATTACTGTTCAAGAGGAGGTTACTACTTTGACTACTGGGGCCAAGGCA

CCACTCTCACAGTCTCCGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTC

CAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACC

GGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTA

CAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGA

AGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGT

CCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTT

CCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTG

GTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAG

GTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGC

GTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAAC

AAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCA

CAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGC

CTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAG

AACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGC

TAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGG

CTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

VAVISSGGSSTFYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAVYYCSRGGYYFDYWGQGTTLT

VSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS

LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLM

ISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN

GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

CCAGATAACCCAGTCTCCATCTTATCTTGCTGCATCTCCTGGAGAAACCATTACTATTAATTGCA

GGGCAAGCAAGACCATTAGCAAATATTTAGCCTGGTATCAAGAGAAACCTGAGAAAACTGATA

AGCTTCTTATCTACTCTGGATCCACTTTGCAATCTGGAATTCCATCAAGGTTCAGTGGCAGTGGA

TCTGGTACAGATTTCACTCTCACCATCAGTGGCCTGGAGCCTGAAGATTTTGCAATGTATTACTG

TCAACAGCATAATGAATACCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAAC

TGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCT

CTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAA

CGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTA

CAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTG

CGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

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

YSGSTLQSGIPSRFSGSGSGTDFTLTISGLEPEDFAMYYCQQHNEYPWTFGGGTKLEIKRTVAAPSVFI

FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA

DYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTGCAAGGCT

TCTGGTTACTCATTCACTGGCTACTACATGCACTGGGTGAAGCAAAGCCATGTAAAGAGCCTTG

AGTGGATTGGACGTATTAATCCTTACAATGGTGCTACTAGCTACAACCAGAATTTCAAGGACAA

GGCCAGCTTGACTGTAGATAAGTCCTCCAGCACAGCCTACATGGAGCTCCACAGCCTGACATCT

GAGGACTCTGCAGTCTATTACTGTGCAAGAGAGGACTACGTCTACTGGGGCCAAGGCACCACTC

TCACAGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAG

CACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACG

GTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCT

CAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTA

CACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATA

TGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTC

CCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGG

ACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATA

ATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCA

CCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCC

TCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGT

ACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA

AAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACT

ACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGT

GGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCA

CAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

WIGRINPYNGATSYNQNFKDKASLTVDKSSSTAYMELHSLTSEDSAVYYCAREDYVYWGQGTTLT

VSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL

SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMI

SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN

GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

GATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGAT

CTAGTCAGAGCCTTGTACACAGTAATGGAAACACCTATTTACATTGGTACCTGCAGAAGCCAGG

CCAGTCTCCAAAGCTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGACAGGTTC

AGTGGCAGTGGATCAGGGACAGATTTCGCACTCAAGATCAGTAGAGTGGAGGCTGAGGATCTG

GGAGTTTATTTCTGCTCTCAAAGTACACATGTTCCGTGGACGTTCGGTGGAGGCACCAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

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

SPKLLIYKVSNRFSGVPDRFSGSGSGTDFALKISRVEAEDLGVYFCSQSTHVPWTFGGGTKLEIKRTV

AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS

TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCT

TCTGGATACACATTCACTGACTATGTTTTGCACTGGGTGAAACAGAAGCCTGGGCAGGGCCTTG

AGTGGATTGGATATATTAATCCTTACAATGATGGTACTAAGTACAATGAGAAGTTCAAAGGCAA

GGCCACACTGACTTCAGACAAATCCTCCAGCACAGCCTACATGGAGCTCAGCAGCCTGACCTCT

GAGGACTCTGCGGTCTATTACTGTGCAAGGGGCTATCCGGCCTACTCTGGGTATGCTATGGACT

ACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCC

CCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGAC

TACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCT

TCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC

AGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGAC

AAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGG

GACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGA

GGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGT

GGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA

CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGC

AAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAG

CCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTC

AGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT

GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC

TCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCG

TGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC

TAGCTGA

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

WIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARGYPAYSGYAMDYW

GQGTSVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL

QSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPP

KPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH

QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDI

AVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS

LSLGKAS

Anti-CD40_12B4.2C10_K-LV-v2-hIgGK-C (SEQ ID NO: 32):
ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGGTACCAGATGTGATATC

CAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCA

GGGCAAGTCAGGACATTAGCAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTA

AACTCCTGATCTACTACACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGG

GTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTT

GCCATCATGGTAATACGCTTCCGTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAA

CTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC

TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATA

ACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCT

ACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCT

GCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTT

AG

Anti-CD40_12B4.2C10_K-LV-v2-hIgGK-C (SEQ ID NO: 97):
MMSSAQFLGLLLLCFQGTRCDIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLL

IYYTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCHHGNTLPWTFGGGTKLEIKRTVAAPSVF

IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGGTGGAGTCTGGGGGAGGCTTAGTGCAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAACC

TCTGGATTCACTTTCAGTGACTATTACATGTATTGGGTTCGCCAGACTCCAGAGAAGAGGCTGG

AGTGGGTCGCATACATTAATTCTGGTGGTGGTAGCACCTATTATCCAGACACTGTAAAGGGCCG

ATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTGCAAATGAGCCGGCTGAAGTCT

GAGGACACAGCCATGTATTACTGTGCAAGACGGGGGTTACCGTTCCATGCTATGGACTATTGGG

GTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGC

GCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTC

CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGG

CTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTG

GGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGA

GTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCAT

CAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCAC

GTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGG

CGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT

GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGT

CTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCG

AGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCT

GACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCA

GCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTAC

AGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATG

CATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCT

GA

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

WVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMSRLKSEDTAMYYCARRGLPFHAMDYWG

QGTSVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ

SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK

PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ

DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGKAS

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

CAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTAGGAGACAGAGTCACCATCAGTTGCA

GTGCAAGTCAGGGCATTAGCAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAA

ACTCCTGATCTATTACACATCAATTTTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGG

TCTGGGACAGATTATTCTCTCACCATCGGCAACCTGGAACCTGAAGATATTGCCACTTACTATTG

TCAGCAGTTTAATAAGCTTCCTCCGACGTTCGGTGGAGGCACCAAACTCGAGATCAAACGAACT

GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTC

TGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAAC

GCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTAC

AGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGC

GAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

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

IYYTSILHSGVPSRFSGSGSGTDYSLTIGNLEPEDIATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFI

FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA

DYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AGTTGGTGCAGTCTGGACCTGAGCTGAAGAAGCCTGGAGAGACAGTCAAGATCTCCTGCAAGG

CTTCTGGGTATTCCTTCACAAACTATGGAATGAACTGGGTGAAACAGGCTCCAGGAAAGGGTTT

AAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGTCAACATATGCTGATGACTTCAAGGG

ACGGTTTGCCTTCTCTTTGGAAACCTCTGCCAGCACTGCCTATTTGCAGATCAGTAACCTCAAAA

ATGAGGACATGGCTACATATTTCTGTGCTAGAGGGGACTTTAGGTACTACTATTTTGACTACTG

GGGCCAAGGCACCACTCTCACAGGCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTG

GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACT

TCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCC

GGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGC

TTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAG

AGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGAC

CATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTC

ACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT

GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGT

GTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAG

GTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCC

GAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCC

TGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC

AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTA

CAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGAT

GCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGC

TGAT

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

KWMGWINTYTGESTYADDFKGRFAFSLETSASTAYLQISNLKNEDMATYFCARGDFRYYYFDYWG

QGTTLTGSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ

SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK

PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ

DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGKAS

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

TCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACGTGT

CGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTC

AGCTCCTGGTCTATAATGCAAAAACCTTGGCAGATGGTGTGCCATCAAGGTTCAGTGGCAGTGG

ATCAGGAACACAATATTCTCTCAAGATCAACACCCTGCAGCCTGAAGATTTTGGGAGTTATTAC

TGTCAACATTTTTGGGATTCTTGGACGTTCGGTGGAGGCACCAAGCTCGAGATCAAACGAACTG

TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCT

GTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACG

CCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACA

GCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGCG

AAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

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

LVYNAKTLADGVPSRFSGSGSGTQYSLKINTLQPEDFGSYYCQHFWDSWTFGGGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAGCAGTCTGGAGCTGAGCTGATGAAGCCTGGGGCCTCAGTGAAGATATCCTGCAAGGC

TACTGGCTACACATTCAGTAGCTACTGGATAGAGTGGGTAAAGCAGAGGCCTGGACATGGCCTT

GAGTGGATTGGAGAGATTTTACCTGGAAGTGGTAGGACTAACGACAATGAGAAGTTCAAGGGC

AAGGCCACATTCACTGCAGATACATCCTCCAAGAAAGCCTACATGCAACTCAGCAGCCTGACAT

CTGAGGACTCTGCCGTCTATTATTGTGCAAGAAGGGGTGGTTACTCCTTTGCTTACTGGGGCCA

AGGGACTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCC

TGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCG

AACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGT

CCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGC

ACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTT

GAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAG

TCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGC

GTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG

GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTC

AGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCC

AACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAG

CCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACC

TGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCG

GAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCA

GGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATG

AGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

WIGEILPGSGRTNDNEKFKGKATFTADTSSKKAYMQLSSLTSEDSAVYYCARRGGYSFAYWGQGTL

VTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

GAAACAACTGTGACCCAGTCTATGACCATGTTCTCACTAGCTCTTCTCCTCAGTCTTCTTCTCCT

CTGTGTCTCTGATTCTAGGGCAGAAACAACTGTGACCCAGTCTCCAGCATCCCTGTCCATGGCT

ATAGGGGAAAAAGTCACCATCAGATGCGTAACCAGCACTGATATTGATGATGATGTGAACTGG

TACCAGCAGAAGCCAGGGGAACCTCCTAAACTCCTTATTTCAGAAGGCAATACTCTTCGTCCTG

GAGTCCCATCCCGATTCTCCAGCAGTGGCTATGGTACAGATTTTGTTTTTACAATTGAGAACATG

CTCTCAGAAGATGTTGCAGATTACTACTGTTTGCAAAGTGGTAACTTGCCGTACACGTTCGGAG

GGGGGACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATC

TGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGA

GAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC

ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCA

GACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTC

ACAAAGAGCTTCAACAGGGGAGAGTGTTAGCCAGCATCCCTGTCCATGGCTATAGGGGAAAAA

GTCACCATCAGATGCGTAACCAGCACTGATATTGATGATGATGTGAACTGGTACCAGCAGAAGC

CAGGGGAACCTCCTAAACTCCTTATTTCAGAAGGCAATACTCTTCGTCCTGGAGTCCCATCCCG

ATTCTCCAGCAGTGGCTATGGTACAGATTTTGTTTTTACAATTGAGAACATGCTCTCAGAAGATG

TTGCAGATTACTACTGTTTGCAAAGTGGTAACTTGCCGTACACGTTCGGAGGGGGGACCAAGCT

CGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTG

AAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTAC

AGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACA

GCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAAC

ACAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAA

CAGGGGAGAGTGTTAG

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

LLISEGNTLRPGVPSRFSSSGYGTDFVFTIENMLSEDVADYYCLQSGNLPYTFGGGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AGTTGGTGCAGTCTGGACCTGAGCTGAAGAAGCCTGGAGAGACAGTCAAGATCTCCTGCAAGG

CTTCTGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGAAAGGGTTT

AAAGTGGGTGGGCTGGATAAACACCTTCACTGGAGAGCCAACATATGTTGATGACTTCAAGGG

ACGGTTTGCCTTCTCTTTGGAAACCTCTGCCAGCACTGCCTATTTGCAGATCAACAACCTCAAAA

ATGAGGACACGGCTACATATTTCTGTGCAAGAGGGAATTTTAGGTACTACTACTTTGACTACTG

GGGCCAAGGCACCACTCTCACAGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTG

GCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACT

TCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCC

GGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGC

TTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAG

AGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGAC

CATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTC

ACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGAT

GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGT

GTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAG

GTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCC

GAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCC

TGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC

AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTA

CAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGAT

GCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGC

TGA

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

KWVGWINTFTGEPTYVDDFKGRFAFSLETSASTAYLQINNLKNEDTATYFCARGNFRYYYFDYWG

QGTTLTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ

SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK

PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ

DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGKAS

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

TCCAGATGACTCAGTCCCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATG

TCGAACAAGTGGGAATATTCGCAATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCT

CAACTCCTGGTCTATAATGCAAAAACCTTAGCAGATGGTGTGCCATCAAGGTTCGGTGGCAGTG

GATCAGGAACACAATATTCTCTCAAGATCAACAGCCTGCAGCCTGAAGATTTTGGGAATTATTA

CTGTCAACATTTTTGGAGTAGTCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACGA

ACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGC

CTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGAT

AACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACC

TACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCC

TGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

TAG

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

LVYNAKTLADGVPSRFGGSGSGTQYSLKINSLQPEDFGNYYCQHFWSSPYTFGGGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

TCCAACTGCAGCAGCCTGGGGCTGAACTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAA

GGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGAGAAGG

CCTTGAGTGGATTGGAGAGATTAATCCTAGCTACGGTCGTACTGACTACAATGAGAAGTTCAAG

AACAAGGCCACACTGACTGTAGCCAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTG

ACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGAGATTACTACGGTAGTAGCTCGTTTG

CTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTT

CCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAG

GACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA

CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCC

AGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTG

GACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAG

GGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCC

TGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTA

CGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC

GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAA

GTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGG

GCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC

AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCT

TCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA

GCTAGCGGATGGAGCTATATCATCCTCTTTTTGGTAGCAACAGCTACAGATGTCCACTCCCAGG

TCCAACTGCAGCAGCCTGGGGCTGAACTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAA

GGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGAGAAGG

CCTTGAGTGGATTGGAGAGATTAATCCTAGCTACGGTCGTACTGACTACAATGGGAAGTTCAAG

AACAAGGCCACACTGACTGTAGCCAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTG

ACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGAGATTACTACGGTAGTAGCTCGTTTG

CTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTT

CCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAG

GACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACA

CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCC

AGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTG

GACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAG

GGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCC

TGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTA

CGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCAC

GTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAA

GTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGG

GCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC

AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCT

TCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA

GCTAGCTGA

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

LEWIGEINPSYGRTDYNEKFKNKATLTVAKSSSTAYMQLSSLTSEDSAVYYCARGDYYGSSSFAYW

GQGTLVTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL

QSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPP

KPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH

QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDI

AVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS

LSLGKAS

Anti-DCIR_29G10.3D9_K-Var1-V-hIgGK-C (SEQ ID NO: 42):
ATGGATTTTCAAGTGCAGATTTTCAGCTTCCTGCTAATGAGTGCCTCAGTCATAATGTCCAGGGG

ACAAATTGTTCTCACCCAGTCTCCAGCACTCATGTCTGCATCTCCAGGGGAGAAGGTCACCATG

ACCTGCAGTGCCAGCTCAAATATAAGTTACATGTACTGGTACCAGCAGAAGCCAAGATCCTCCC

CCAAACCCTGGATTTATCTCACATCCAACCTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGT

GGGTCTGGGACCTCTTACTCTCTCACAACCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATT

GCTGCCAGCAGTGGAGTAGTAACCCACCCACGTTCGGTGCTGGGACCAAGCTCGAGATCAAAC

GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT

GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGG

ATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCA

CCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATG

CCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT

GTTAG

Anti-DCIR_29G10.3D9_K-Var1-V-hIgGK-C (SEQ ID NO: 107):
MDFQVQIFSFLLMSASVIMSRGQIVLTQSPALMSASPGEKVTMTCSASSNISYMYWYQQKPRSSPKP

WIYLTSNLASGVPARFSGSGSGTSYSLTTSSMEAEDAATYCCQQWSSNPPTFGAGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Anti-DCIR_29G10.3D9_K-Var2-V-hIgGK-C (SEQ ID NO: 43):
ATGGATTTTCGAGTGCAGATTTTCAGCTTCCTGCTAATGAGTGCCTCAGTCATAATGTCCAGGGG

ACAAATTGTTCTCACCCAGTCTCCAGCACTCATGTCTGCATCTCCAGGGGAGAAGGTCACCATG

ACCTGCAGTGCCAGCTCAAATATAAGTTACATGTACTGGTACCAGCAGAAGCCAAGATCCTCCC

CCAAACCCTGGATTTATCTCACATCCAACCTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGT

GGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATT

ACTGCCAGCAGTGGAGTAGTAACCCACCCACGTTCGGTGCTGGGACCAAGCTCGAGATCAAAC

GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT

GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGG

ATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCA

CCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATG

CCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT

GTTAG

Anti-DCIR_29G10.3D9_K-Var2-V-hIgGK-C (SEQ ID NO: 108):
MDFRVQIFSFLLMSASVIMSRGQIVLTQSPALMSASPGEKVTMTCSASSNISYMYWYQQKPRSSPKP

WIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGAGTKLEIKRTVAAPS

VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL

SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

TTGTGCTGATCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGC

AGAGCCAGTGAAAGTGTTGATAGTTATGTCAATAGTTTTATGCACTGGTACCAGCAGAAACCAG

GACAGCCACCCAAACTCCTCATCTATCGTGTATCCAACCTAGAATCTGGGATCCCTGCCAGGTT

CAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTAATCCTGTGGAGGCTGATGATGTT

GCAACCTATTACTGTCAGCAAAGTAATGAGGATCCATTCACGTTCGGCTCGGGGACAAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

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

PPKLLIYRVSNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPFTFGSGTKLEIKRTVA

APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST

LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

Anti-DCIR_31A6.1F5_H-var2-V-hIgG4H-C (SEQ ID NO: 45):
ATGGAATGTAACTGGATACTTCCTTTTATTCTGTCGGTAATTTCAGGGGTCTACTCAGAGGTTCA

GCTCCAGCAGTCTGGGACTGTGCTGGCAAGGCCTGGGGCTTCCGTGAATATGTCCTGTAAGGCT

GCTGGCTACAGCTTTACCAGTTACTGGGTGTACTGGGTCAAACAGAGGCCTGGACAGGGTCTGG

AATGGATTGGTGCTATTTACCCTAAAAATAGTAGAACTAGCTACAACCAGAAGTTCCAGGACAA

GGCCACACTGACTGCAGTCACATCCGCCAGCACTGCCTACATGGAGCTCAGCAGCCTGACAAAT

GAGGACTCTGCGGTCTATTACTGTACAAGACCTCACTATGATTCGTTTGGTTACTGGGGCCAAG

GGACTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTG

CTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAA

CCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCC

TACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCAC

GAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGA

GTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTC

TTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCG

TGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGG

AGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCA

GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCA

ACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGC

CACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCT

GCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG

AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAG

GCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGA

GGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

Anti-DCIR_31A6.1F5_H-var2-V-hIgG4H-C (SEQ ID NO: 110):
MECNWILPFILSVISGVYSEVQLQQSGTVLARPGASVNMSCKAAGYSFTSYWVYWVKQRPGQGLE

WIGAIYPKNSRTSYNQKFQDKATLTAVTSASTAYMELSSLTNEDSAVYYCTRPHYDSFGYWGQGTL

VTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

Anti-DCIR_31A6.1F5_K-var2-V-hIgGK-C (SEQ ID NO: 46):
ATGGAGACAGACACACTCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGGTTCCACAGGTGACA

TTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGC

AGAGCCAGTGAAAGTGTAGATAGTTATGGCATTAGTTTTATGCACTGGTACCAGCAGAAACCAG

GACAGCCACCCAAACTCCTCATCTATCGTGCATCCAACCAAGAATCTGGGATCCCTGCCAGGTT

CAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTAATCCTGTGGAGGCTGATGATGTT

GCAACCTATTACTGTCAGCAAAGTAATGAGGATCCGCTCACGTTCGGTGCTGGGACCAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

Anti-DCIR_31A6.1F5_K-var2-V-hIgGK-C (SEQ ID NO: 111):
METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISCRASESVDSYGISFMHWYQQKPGQ

PPKLLIYRASNQESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPLTFGAGTKLEIKRTVA

APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST

LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

TCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCTTTCT

CTGGGTTTTCACTGAGCACTTCTGGTATGGGTGTGAGCTGGATTCGTCAGCCTTCAGGAAAGGG

TCTGGAGTGGCTGGCACACATTTACTGGGATGATGACAAGCGCTATAATCCATCCCTGAAGAGC

CGGCTCACAATCTTTAAGGATCCCTCCAGCAACCAGGTATTCCTCAGGATCACCAGTGTGGACA

CTGCAGATACTGCCACATACTACTGTGCTCGAAACTCCCATTACTACGGTAGTACTTACGGGGG

ATACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCAGCCAAAACAAAGGGCCC

ATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGC

CTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCG

GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGAC

CGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAA

CACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT

GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCT

CCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGT

TCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGT

TCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAA

GGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAA

AGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGAC

CAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG

TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCT

TCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTC

TCTGGGTAAAGCTAGCTGA

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

AHIYWDDDKRYNPSLKSRLTIFKDPSSNQVFLRITSVDTADTATYYCARNSHYYGSTYGGYFDVWG

AGTTVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ

SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK

PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ

DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGKAS.

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

TTGTGCTGACCCAGTCTCCAACTTCTTTCACTGTGTCTCTTGGGCAGAGGGCCACCATATCCTGC

AGAGCCAGTGAAAGTGTTCATAGTTATGGCAATAGTTTTATGCACTGGTACCAGCAGAAACCAG

GGCAGCCACCCAAACTCCTCATCTATCTTGCATCCAACGTAGAATCTGGGGTCCCTGCCAGGTT

CAGTGGTAGTGGGTCCAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCT

GCAACCTATTACTGTCAGCAAAATAGTGAGGATCCGTGGACGTTCGGTGGAGGCACCAAGCTC

GAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGA

AATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA

GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAG

CAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACA

CAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC

AGGGGAGAGTGTTAG

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

PKLLIYLASNVESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNSEDPWTFGGGTKLEIKRTV

AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS

TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAGCAGTCTGGAACTGAGCTGATGAAGCCTGGGGCCTCAGTGAAGATATCCTGCAAGGC

TACTGGCTACACATTCAGTACCTACTGGATAGAGTGGGTAAAGCAGAGGCCTGGACATGGCCTT

GAGTGGATTGGAGAGATTTTACCTGGAAGTGGTAGGACTAACGACAATGAGAAGTTCAAGGGC

AAGGCCACAATCACTGCAGATACATCCTCCAAGAAAGCCTACATGCAACTCAGCAGCCTGACA

TCTGAGGACTCTGCCGTCTATTACTGTGCAAGAAGGGGTGGTTACTCCTTTGCTTTCTGGGGCCA

AGGGACTCTGGTCTCTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCCCTGGCGCCC

TGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCG

AACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGT

CCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGC

ACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTT

GAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAG

TCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGC

GTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTG

GAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTC

AGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCC

AACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAG

CCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACC

TGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCG

GAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCA

GGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATG

AGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

WIGEILPGSGRTNDNEKFKGKATITADTSSKKAYMQLSSLTSEDSAVYYCARRGGYSFAFWGQGTL

VSVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

GAAACAACTGTGACCCAGTCTCCAGCATCCCTGTCCATGGCTATAGGAGAAAAAGTCACCATCA

GATGCGTAACCAGCACTGATATTGATGATGATGTGAACTGGTACCAGCAGAAGCCAGGGGAAC

CTCCTAAGCTCCTTATTTCAGAAGGCAATACTCTTCGTGCTGGAGTCCCATCCCGATTCTCCAGC

AGTGGCTATGGTACAGATTTTGTTTTTACAATTGAGAACATGCTCTCAGAAGATGTTGCAGATT

ACTACTGTTTGCAAAGTGGTAACTTGCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAA

ACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGA

ACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGG

TGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACA

GCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCT

ATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG

AGTGTTAG

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

LLISEGNTLRAGVPSRFSSSGYGTDFVFTIENMLSEDVADYYCLQSGNLPYTFGGGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT

LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

TCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCTTTCT

CTGGGTTTTCACTGAGCACTTCTGGTATGGGTCTGAGCTGGATTCGTCAGCCTTCAGGAAAGGG

TCTGGAGTGGCTGGCACACATTTACTGGGATGATGACAAGCGCTATAACCCATCCCTGAAGAGC

CGGCTCACAATCTCCAAGGATACCTCCAGCAACCAGGTTTTCCTCAAGATCACCATTGTGGACA

CTGCAGATGCTGCCACATACTACTGTGCTCGAAGCTCCCATTACTACGGTTATGGCTACGGGGG

ATACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCAGCCAAAACGAAGGGCCC

ATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGC

CTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCG

GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGAC

CGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAA

CACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCT

GAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCT

CCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGT

TCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGT

TCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAA

GGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAA

AGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGAC

CAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAG

TGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCT

TCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTC

TCTGGGTAAAGCTAGCTGA

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

AHIYWDDDKRYNPSLKSRLTISKDTSSNQVFLKITIVDTADAATYYCARSSHYYGYGYGGYFDVWG

AGTTVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ

SSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK

PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ

DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGKAS

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

TTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGC

AGAGCCAGTGAAAGTATTCATAGTTATGGCAATAGTTTTCTGCACTGGTACCAGCAGAAACCAG

GACAGCCACCCAAACTCCTCATCTATCTTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTT

CAGCGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCT

GCAACCTATTACTGTCAGCAAAATAATGAGGATCCGTGGACGTTCGGTGGAGGCACCAAGCTC

GAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGA

AATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA

GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAG

CAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACA

CAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC

AGGGGAGAGTGTTAGGCGGCCGCACTAGCGCGGGCCGCATTCGAAGAGCTCGGTACCCGGGGA

TCCTCTAGAGTCGACCTGCAGGCATGCAAGCTGGCCGCGACTCTAGATCATAATCAGC

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

PKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPWTFGGGTKLEIKRTV

AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS

TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AGCTGCAGCAGTCTGGGGCTGAGCTTGTGAGGCCAGGGGCCTTAGTCAAGTTGTCCTGCAAAGC

TTCTGGCTTCAACATTAATGACTACTATATCCACTGGGTGAAGCAGCGGCCTGAACAGGGCCTG

GAGCGGATTGGATGGATTGATCCTGACAATGGTAATACTATATATGACCCGAAGTTCCAGGGCA

AGGCCAGTATAACAGCAGACACATCCCCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACAT

CTGAGGACACTGCCGTCTATTACTGTGCTAGAACCCGATCTCCTATGGTTACGACGGGGTTTGTT

TACTGGGGCCAAGGGACTGTGGTCACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCC

CCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGA

CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACC

TTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAG

CAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGA

CAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGG

GGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTG

AGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACG

TGGATGGCGTGGAGGTGCATAATGCCAAGACRAAGCCGCGGGAGGAGCAGTTCAACAGCACGT

ACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGT

GCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGC

AGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGG

TCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAA

TGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTC

CTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCC

GTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAT

GA

Anti-DCIR2C9H-LV-hIgG4H-V-hIgG4H-C (SEQ ID NO: 118):
MKCSWVIFFLMAVVTGVNSEVQLQQSGAELVRPGALVKLSCKASGFNINDYYIHWVKQRPEQGLE

RIGWIDPDNGNTIYDPKFQGKASITADTSPNTAYLQLSSLTSEDTAVYYCARTRSPMVTTGFVYWGQ

GTVVTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS

SGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKP

KDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKXKPREEQFNSTYRVVSVLTVLHQD

WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAV

EWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL

GK

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

GCTTCAGCAGTCTGGGGCTGAGCTGGCAAAACCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCT

TCTGGCTACACCTTTACTACCTACTGGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGG

AATGGATTGGATACATTAATCCTATCACTGGTTATACTGAGTACAATCAGAAGTTCAAGGACAA

GGCCACCTTGACTGCAGACAAATCCTCCAGCACAGCCTACATGCAACTGAGCAGCCTGACATCT

GAGGACTCTGCAGTCTATTACTGTGCAAGAGAGGGTTTAAGTGCTATGGACTATTGGGGTCAGG

GAACCTCAGTCACCGTCACCTCAGCCAAAACAACAGCCCCATCGGTCTATCCACTGGCCCCTGT

GTGTGGAGATACAACTGGCTCCTCGGTAACTCTAGGATGCCTGGTCAAGGGTTATTTCCCTGAG

CCAGTGACCTTGACCTGGAACTCTGGATCCCTGTCCAGTGGTGTGCACACCTTCCCAGCTGTCCT

GCAGTCTGACCTCTACACCCTCAGCAGCTCAGTGACTGTAACCTCGAGCACCTGGCCCAGCCAG

ACCGTCACCTGCAGCGTTGCTCACCCAGCCAGCAGCACCACGGTGGACAAAAAACTTGAGCCC

AGCGGGCCCATTTCAACAATCAACCCCTGTCCTCCATGCAAGGAGTGTCACAAATGCCCAGCTC

CTAACCTCGAGGGTGGACCATCCGTCTTCATCTTCCCTCCAAATATCAAGGATGTACTCATGATC

TCCCTGACACCCAAGGTCACGTGTGTGGTGGTGGATGTGAGCGAGGATGACCCAGACGTCCAG

ATCAGCTGGTTTGTGAACAACGTGGAAGTACACACAGCTCAGACACAAACCCATAGAGAGGAT

TACAACAGTACTATCCGGGTGGTCAGCACCCTCCCCATCCAGCACCAGGACTGGATGAGTGGCA

AGGAGTTCAAATGCAAGGTCAACAACAAAGACCTCCCATCACCCATCGAGAGAACCATCTCAA

AAATTAAAGGGCTAGTCAGAGCTCCACAAGTATACATCTTGCCGCCACCAGCAGAGCAGTTGTC

CAGGAAAGATGTCAGTCTCACTTGCCTGGTCGTGGGCTTCAACCCTGGAGACATCAGTGTGGAG

TGGACCAGCAATGGGCATACAGAGGAGAACTACAAGGACACCGCACCAGTCCTGGACTCTGAC

GGTTCTTACTTCATATATAGCAAGCTCAATATGAAAACAAGCAAGTGGGAGAAAACAGATTCCT

TCTCATGCAACGTGAGACACGAGGGTCTGAAAAATTACTACCTGAAGAAGACCATCTCCCGGTC

TCCGGGTAAAGCTAGCTGA

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

EWIGYINPITGYTEYNQKFKDKATLTADKSSSTAYMQLSSLTSEDSAVYYCAREGLSAMDYWGQGT

SVTVTSAKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDL

YTLSSSVTVTSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPCKECHKCPAPNLEGGPSVFI

FPPNIKDVLMISLTPKVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTIRVVSTLPIQ

HQDWMSGKEFKCKVNNKDLPSPIERTISKIKGLVRAPQVYILPPPAEQLSRKDVSLTCLVVGFNPGDI

SVEWTSNGHTEENYKDTAPVLDSDGSYFIYSKLNMKTSKWEKTDSFSCNVRHEGLKNYYLKKTISR

SPGKAS

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

TGGCGGAGACATTGTGATGACCCAGTCTCACAAATTCATGTCCACATCAGTAGGAGACAGGGTC

AGCGTCACCTGCAAGGCCAGTCAGGATGTGACTTCTGCTGTAGCCTGGTATCAACAAAAACCAG

GGCAATCTCCTAAACTACTGATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCTT

CACAGGCAGTGGATCTGGGACAGATTATACTCTCACCATCAGCAGTGGGCAGGCTGAAGACCT

GGCACTTTATTACTGTCACCAATATTATAGCGCTCCTCGGACGTTCGGTGGAGGCACCAAGCTG

GAAGTCAAACGGGCTGATGCTGCACCAACTGTATCCATCTTCCCACCATCCAGTGAGCAGTTAA

CATCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTACCCCAAAGACATCAATGTCAA

GTGGAAGATTGATGGCAGTGAACGACAAAATGGCGTCCTGAACAGTTGGACTGATCAGGACAG

CAAAGACAGCACCTACAGCATGAGCAGCACCCTCACGTTGACCAAGGACGAGTATGAACGACA

TAACAGCTATACCTGTGAGGCCACTCACAAGACATCAACTTCACCCATCGTCAAGAGCTTCAAT

AGGAATGAGTGTTAG

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

LLIYWASTRHTGVPDRFTGSGSGTDYTLTISSGQAEDLALYYCHQYYSAPRTFGGGTKLEVKRADA

APTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSST

LTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

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

GCTTCAGCAGTCTGGGGCTGAGCTGGCAAAACCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCT

TCTGGCTACACCTTTACTACCTACTGGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGG

AATGGATTGGATACATTAATCCTATCACTGGTTATACTGAGTACAATCAGAAGTTCAAGGACAA

GGCCACCTTGACTGCAGACAAATCCTCCAGCACAGCCTACATGCAACTGAGCAGCCTGACATCT

GAGGACTCTGCAGTCTATTACTGTGCAAGAGAGGGTTTAAGTGCTATGGACTATTGGGGTCAGG

GAACCTCAGTCACCGTCACCTCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCGCCCTG

CTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAA

CCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCC

TACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCAC

GAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGA

GTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTC

TTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCG

TGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGG

AGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCA

GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCA

ACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGC

CACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCT

GCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG

AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAG

GCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGA

GGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

EWIGYINPITGYTEYNQKFKDKATLTADKSSSTAYMQLSSLTSEDSAVYYCAREGLSAMDYWGQGT

SVTVTSAKTTGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKA

S.

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

TGGCGGAGACATTGTGATGACCCAGTCTCACAAATTCATGTCCACATCAGTAGGAGACAGGGTC

AGCGTCACCTGCAAGGCCAGTCAGGATGTGACTTCTGCTGTAGCCTGGTATCAACAAAAACCAG

GGCAATCTCCTAAACTACTGATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCTT

CACAGGCAGTGGATCTGGGACAGATTATACTCTCACCATCAGCAGTGGGCAGGCTGAAGACCT

GGCACTTTATTACTGTCACCAATATTATAGCGCTCCTCGGACGTTCGGTGGAGGCACCAAGCTC

GAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGA

AATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA

GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAG

CAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACA

CAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC

AGGGGAGAGTGTTAG

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

LLIYWASTRHTGVPDRFTGSGSGTDYTLTISSGQAEDLALYYCHQYYSAPRTFGGGTKLEIKRTVAA

PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL

TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

ACTGAAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCATTACCTGCTCTGTC

TCTGGGTTCTCATTAAGCAACTATGATATAAGCTGGATTCGCCAGCCACCAGGAAAGGGTCTGG

AGTGGCTTGGAGTAATGTGGACTGGTGGAGGCGCAAATTATAATTCAGCTTTCATGTCCAGACT

GAGCATCAACAAGGACAACTCCAAGAGCCAAGTTTTTTTAAAAATGAACAATCTGCAAACTGA

TGACACAGCCATTTATTACTGTGTCAGAGATGCGGTGAGGTACTGGAACTTCGATGTCTGGGGC

GCAGGGACCACGGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGC

CCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCC

CGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCT

GTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGG

GCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAG

TTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATC

AGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACG

TGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGC

GTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTG

GTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTC

TCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGA

GAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTG

ACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAG

CCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACA

GCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGC

ATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

Anti-Dectin_1_11B6.4_H-V-hIgG4H-C (SEQ ID NO: 123):
MAVLALLLCLVAFPTCTLSQVQLKESGPGLVAPSQSLSITCSVSGFSLSNYDISWIRQPPGKGLEWLG

VMWTGGGANYNSAFMSRLSINKDNSKSQVFLKMNNLQTDDTAIYYCVRDAVRYWNFDVWGAGT

TVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

ACAAATTGTTCTCTCCCAGTCACCAGCAATCCTGTCTGCATCTCCAGGGGAGAAGGTCACAATG

ACTTGCAGGGCCAGCTCAAGTGTAAGTTACATACACTGGTACCAGCAGAAGCCAGGATCCTCCC

CCAAACCCTGGATTTATGCCACATCCCACCTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGT

GGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGAGTGGAGGCTGAAGATACTGCCACTTATT

ACTGCCAGCAGTGGAGTAGTAACCCATTCACGTTCGGCTCGGGGACAAAGCTCGAGATCAAAC

GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT

GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGG

ATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCA

CCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATG

CCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT

GTTAG

Anti-Dectin_1_11B6.4_K-LV-hIgGK-C (SEQ ID NO: 124):
MDFQAQIFSFLLISASVIMSRGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWYQQKPGSSPKPWI

YATSHLASGVPARFSGSGSGTSYSLTISRVEAEDTATYYCQQWSSNPFTFGSGTKLEIKRTVAAPSVF

IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AGCTGCAGCAGTCTGGGGCTGAACTGGCAAGACCTGGGGCCTCAGTGAAGATGTCCTGCAAGG

CTTCTGGCTACACCTTTACTACCTACACTATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCT

GGAATGGATTGGATACATTAATCCTAGCAGTGGTTATACTAATTACAATCAGAAGTTCAAGGAC

AAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGCCTCCATGCAACTGAGCAGCCTGACAT

CTGAGGACTCTGCAGTCTATTACTGTGCAAGAGAGAGGGCGGTATTAGTCCCCTATGCTATGGA

CTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACAAAGGGCCCATCCGTCTTC

CCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGG

ACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACAC

CTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCA

GCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGG

ACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGG

GGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCT

GAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTAC

GTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACG

TACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGT

GCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGC

AGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGG

TCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAA

TGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTC

CTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCC

GTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAG

CTAGCTGA

Anti-Dectin_1_15E2.5_H-V-hIgG4H-C (SEQ ID NO: 125):
MERHWIFLLLLSVTAGVHSQVQLQQSGAELARPGASVKMSCKASGYTFTTYTMHWVKQRPGQGL

EWIGYINPSSGYTNYNQKFKDKATLTADKSSSTASMQLSSLTSEDSAVYYCARERAVLVPYAMDY

WGQGTSVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV

LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFP

PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVL

HQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSL

SLSLGKAS

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

ACAAATTGTTCTCACCCAGTCTCCAGCAGTCATGTCTGCATCTCCAGGGGAGAAGGTCACCATA

ACCTGCACTGCCAGCTCAAGTTTAAGTTACATGCACTGGTTCCAGCAGAAGCCAGGCACTTCTC

CCAAACTCTGGCTTTATAGCACATCCATCCTGGCTTCTGGAGTCCCTACTCGCTTCAGTGGCAGT

GGATCTGGGACCTCTTACTCTCTCACAATCAGCCGAATGGAGGCTGAAGATGCTGCCACTTATT

ACTGCCAGCAAAGGAGTAGTTCCCCATTCACGTTCGGCTCGGGGACAAAGCTCGAGATCAAAC

GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACT

GCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGG

ATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCA

CCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATG

CCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT

GTTAG

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

LYSTSILASGVPTRFSGSGSGTSYSLTISRMEAEDAATYYCQQRSSSPFTFGSGTKLEIKRTVAAPSVFI

FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA

DYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAGCAGTCTGGACCTGAGCTGGAGAAGCCTGGCGCTTCAGTGAAGATATCCTGCAAGGCT

TCTGGTTACTCCTTCACTGGCTACAACATGAACTGGGTGAAACAGAGCAATGGAAAGAGCCTTG

AGTGGATTGGAAATATTGATCCTTACTATGGTGATACTAACTACAACCAGAAGTTCAAGGGCAA

GGCCACATTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCACCTCAAGAGCCTGACATCT

GAGGACTCTGCAGTCTATTACTGTGCAAGACCCTACGGTAGTGAGGCCTACTTTGCTTACTGGG

GCCAAGGGACTCTGGTCACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGC

GCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTC

CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGG

CTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTG

GGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGA

GTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCAT

CAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCAC

GTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGG

CGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT

GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGT

CTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCG

AGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCT

GACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCA

GCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTAC

AGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATG

CATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCT

GA

Anti-Dectin_1_2D8.2D4H-V-hIgG4H-C (SEQ ID NO: 127):
MGWTWIFILILSVTTGVHSEVQLQQSGPELEKPGASVKISCKASGYSFTGYNMNWVKQSNGKSLEW

IGNIDPYYGDTNYNQKFKGKATLTVDKSSSTAYMHLKSLTSEDSAVYYCARPYGSEAYFAYWGQG

TLVTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS

GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPK

DTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDW

LNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE

WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

KAS

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

GTGATGACTCAGTCTCCAGCCACCCTGTCTGTGACTCCAGGAGATAGAGTCTCTCTTTCCTGCAG

GGCCAGCCAGAGTATTAGCGACTACTTACACTGGTATCAACAAAAATCACATGAGTCTCCAAGG

CTTCTCATCAAATATGCTGCCCAATCCATCTCTGGGATCCCCTCCAGGTTCAGTGGCAGTGGATC

AGGGTCAGATTTCACTCTCAGTATCAACGGTGTGGAACCTGAAGATGTTGGAGTGTATTACTGT

CAAAATGGTCACAGCTTTCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACGAACT

GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTC

TGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAAC

GCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTAC

AGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGCCTGC

GAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

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

DFTLSINGVEPEDVGVYYCQNGHSFPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN

NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS

SPVTKSFNRGEC

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

GCGGCAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCT

GGATACACATTTACTGACTATGTTATAAGTTGGGTGAAGCAGAGAACTGGACAGGGCCTTGAGT

GGATTGGAGATATTTATCCTGGAAGTGGTTATTCTTTCTACAATGAGAACTTCAAGGGCAAGGC

CACACTGACTGCAGACAAATCCTCCACCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAG

GACTCTGCGGTCTATTTCTGTGCAACCTACTATAACTACCCTTTTGCTTACTGGGGCCAAGGGAC

TCTGGTCACTGTCTCTGCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCA

GGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGT

GACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAG

TCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGA

CCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCA

AATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCT

GTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTG

GTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTG

CATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTC

CTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAA

GGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAG

GTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG

GTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAAC

AACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAA

CCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCT

GCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

KATLTADKSSTTAYMQLSSLTSEDSAVYFCATYYNYPFAYWGQGTLVTVSAAKTTGPSVFPLAPCS

RSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTC

NVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDP

EVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI

SKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG

SFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS.

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

GATGACCCAAACTCCACTCTCCCTGCCTGTCCGTCTTGGAGATCAAGCCTCCATCTCTTGCAGAT

CTAGTCAGAGCCTTGTACACAGTAATGGAAACACCTATTTACATTGGTACCTGCAGAAGCCAGG

CCAGTCTCCAAAGCTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGACAGGTTC

AGTGGCAGTGGATCAGGGACAAATTTCACACTCAAGATCAGCAGAGTGGAGGCTGAGGATCTG

GGACTTTATTTCTGCTCTCAAAGTACACATGTTCCGTACACGTTCGGAGGGGGGACCAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

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

SGSGSGTNFTLKISRVEAEDLGLYFCSQSTHVPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTAS

VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV

THQGLSSPVTKSFNRGEC

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

TTGTGAGGTGCAGCTTGTTGAGTCTGGTGGAGGATTGGTGCAGCCTAAAGGGTCATTGAAACTC

TCATGTGCAGCCTCTGGATTAACCTTCAATATCTACGCCATGAACTGGGTCCGCCAGGCTCCAG

GAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAATAAAAGTAATAATTATGCAACATATTATGC

CGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCACAAAGCTTGCTCTATCTGCAA

ATGAACAACTTGAAAACTGAGGACACAGCCATGTATTACTGTGTGGGACGGGACTGGTTTGATT

ACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCCC

CCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGAC

TACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCT

TCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC

AGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGAC

AAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGG

GACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGA

GGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGT

GGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA

CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGC

AAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAG

CCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTC

AGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT

GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC

TCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCG

TGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC

TAGCTGA

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

GKGLEWVARIRNKSNNYATYYADSVKDRFTISRDDSQSLLYLQMNNLKTEDTAMYYCVGRDWFD

YWGQGTLVTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP

AVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFL

FPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTV

LHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSL

SLSLGKAS

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

TGTGACTCAGGAATCTGCACTCACCACATCACCTGGTGAAACAGTCACACTCACTTGTCGCTCA

AGTACTGGGGCTGTTACAACTAGTAACTATGCCAACTGGGTCCAAGAAAAACCAGATCATTTAT

TCACTGGTCTAATAGGTGGTACCAACAACCGAGTTTCAGGTGTTCCTGCCAGATTCTCAGGCTC

CCTGATTGGAGACAAGGCTGCCCTCACCATCACAGGGGCACAGACTGAGGATGAGGCAATATA

TTTCTGTGCTCTATGGTACAGCAACCATTGGGTGTTCGGTGGAGGAACCAAACTCGAGATCAAA

CGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAAC

TGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTG

GATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGC

ACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAT

GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAG

TGTTAG

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

IGGTNNRVSGVPARFSGSLIGDKAALTITGAQTEDEAIYFCALWYSNHWVFGGGTKLEIKRTVAAPS

VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL

SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGCAGCAGTCTGGAGCTGAGCTGATGAAGCCTGGGGCCTCAGTGAAGATATCCTGCAAGGC

TACTGGCTACACATTCGGTAGCTACTGGATAGAGTGGGTAAAGCAGAGGCCTGGACATGGCCTT

GAGTGGATTGGAGAGATTTTACCTGGAAGTGGTAATACTAACTACAATGAGAACTTCAAGGGC

AAGGCCACATTCACTGCAGATACATCCTCCAACACAGCCTACATGCAACTCACCAGTCTGACAT

CTGAGGACTCTGCCGTCTATTACTGTGCTAGGGCGGGGATTTATTGGGGCCAAGGGACTCTGGT

CACTGTCTCTGCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGC

ACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGG

TGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTC

AGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTAC

ACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATAT

GGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGTTCC

CCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGA

CGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAA

TGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCAC

CGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCT

CCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTA

CACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAA

AGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTA

CAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTG

GACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

WIGEILPGSGNTNYNENFKGKATFTADTSSNTAYMQLTSLTSEDSAVYYCARAGIYWGQGTLVTVS

AAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS

VVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISR

TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEY

KCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ

PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

TTGTGCTGACCCAATCTCCAGCTTTTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCTCCTGC

AGAGCCAGCGAAAGTGTTGATAATTATGGCATTAGTTTTATGAACTGGTTCCAACAGAAACCAG

GACAGCCACCCAAACTCCTCATCTATGTTGCATCCAAGCAAGGATCCGGGGTCCCTGCCAGGTT

TAGTGGCAGTGGGTCTGGGACAGACTTCAGCCTCAACATCCATCCTATGGAGGAGGATGATACT

GCAATGTATTTCTGTCAGCAAAGTAAGGAGGTTCCTCGGACGTTCGGTGGAGGCACCAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

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

PPKLLIYVASKQGSGVPARFSGSGSGTDFSLNIHPMEEDDTAMYFCQQSKEVPRTFGGGTKLEIKRT

VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS

STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTCCAGCAGTCTGGGACTGTGCTGGCAAGGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCT

TCTGGCTACACCTTTACCAGCTACTGGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGG

AATGGATTGGCGCTATTTATCCTGGAAATAGTGATACTACCTACAACCAGAAGTTCAAGGGCAA

GGCCAAACTGACTGCAGTCACATCCACCAGCACTGCCTACATGGAGCTCAGCAGCCTGACAAAT

GAGGACTCTGCGGTCTATTACTGTACACCTACTTACTACTTTGACTACTGGGGCCAAGGCACCTC

TCTCACAGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGG

AGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGA

CGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTC

CTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACC

TACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAA

TATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCATCAGTCTTCCTGT

TCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGT

GGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCA

TAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCT

CACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGG

CCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGT

GTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTC

AAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAAC

TACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCG

TGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGC

ACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTGA

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

WIGAIYPGNSDTTYNQKFKGKAKLTAVTSTSTAYMELSSLTNEDSAVYYCTPTYYFDYWGQGTSLT

VSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL

SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMI

SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGK

EYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN

GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

GATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGT

CAAGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTCTTACAGAGGCCAGG

CCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAACTGGACTCTGGAGTCCCTGACAGGTTC

ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTG

GGAGTTTATTATTGCTGGCAAGGTACACATTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

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

KRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPWTFGGGTKLEIKRTVA

APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST

LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AGCTGCAGCAGACTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTGCAAGG

CTTCTGGTTATCCATTCACTGACTACATCATGGTCTGGGTGAAGCAGAGCCATGGAAAGAGCCT

TGAGTGGATTGGAAATATTAGTCCTTACTATGGTACTACTAACTACAATCTGAAGTTCAAGGGC

AAGGCCACATTGACTGTAGACAAATCTTCCAGCACAGCCTACATGCAGCTCAACAGTCTGACAT

CTGAGGACTCTGCAGTCTATTACTGTGCAAGATCCCCTAACTGGGACGGGGCCTGGTTTGCTCA

CTGGGGCCAAGGGGCTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCC

CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACT

ACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTT

CCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC

AGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGAC

AAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGG

GACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGA

GGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGT

GGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA

CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGC

AAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAG

CCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTC

AGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT

GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC

TCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCG

TGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC

TAGCTGATTAATTAA

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

GNISPYYGTTNYNLKFKGKATLTVDKSSSTAYMQLNSLTSEDSAVYYCARSPNWDGAWFAHWGQ

GALVTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS

SGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKP

KDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQD

WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAV

EWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL

GKAS

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

TTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCTCCTGC

AAGGCCAGCCAAAGTGTTGATTATGATGGTGATAGTTATATGAACTGGTTCCAACAGAAACCAG

GACAGCCACCCAAACTCCTCATCTATGCTGCATCCAATCTAGAATCTGGGATCCCAGCCAGGTT

TAGTGGCAGTGGGTCTGGGACAGACTTCACCCTCAACATCCATCCTGTGGAGGAGGAGGATGCT

GCAACCTATTACTGTCAGCAAAGTAATGAGGATCCATTCACGTTCGGCTCGGGGACAAAGCTCG

AGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA

ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGTTAG

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

PPKLLIYAASNLESGIPARFSGSGSGTDFTLNIHPVEEEDAATYYCQQSNEDPFTFGSGTKLEIKRTVA

APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST

LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GCTGAAGGAGTCAGGACCTGGCCTGGTGGCACCCTCACAGAGCCTGTCCATCACATGCACTGTC

TCTGGGTTCTCATTATCCAGATATAGTGTATTTTGGGTTCGCCAGCCTCCAGGAAAGGGTCTGGA

GTGGCTGGGATTGATATGGGGTGGTGGAAGCACAGACTATAATTCAGCTCTCAAATCCAGACTG

AGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGAT

GACACAGCCATGTACTACTGTGCCAGAATCTACTTTGATTACGACGGGGCTATGGACTACTGGG

GTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACAACGGGCCCATCCGTCTTCCCCCTGGC

GCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTC

CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGG

CTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTG

GGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGA

GTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCAT

CAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCAC

GTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGG

CGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT

GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGT

CTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCG

AGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCT

GACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCA

GCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTAC

AGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATG

CATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCTG

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

GLIWGGGSTDYNSALKSRLSISKDNSKSQVFLKMNSLQTDDTAMYYCARIYFDYDGAMDYWGQGT

SVTVSSAKTTGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL

YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDT

LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE

SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

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

TCTGGTTGTCTGGTGTTGGCGGAGACATTGTGATGACCCAGTCTCACAAATTCATGTCCACATCA

GTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGGATGTGACTTCTGCTGTAGCCTGGT

ATCAACAAAAACCAGGGCAATCTCCTAAACTACTGATTTACTGGGCATCCACCCGGCACACTGG

AGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTATACTCTCACCATCAGCAGTGGG

CAGGCTGAAGACCTGGCACTTTATTACTGTCACCAATATTATAGCGCTCCTCGGACGTTCGGTG

GAGGCACCAAGCTCGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATC

TGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGA

GAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC

ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCA

GACTACGAGAAACACAAAGTCTATGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTC

ACAAAGAGCTTCAACAGGGGAGAGTGTTAG

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

QQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDYTLTISSGQAEDLALYYCHQYYSAPRTFGGGTK

LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD

STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

GATGCAGCAGTCTGGAGCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGACT

TCTGGCTTCACCTTCAGCAGTAACTATATAAGTTGGTTGAAGCAAAAGCCTGGACAGAGTCTTG

AGTGGATTGCATGGATTTATGCTGGAACTGGTGGTATTACCTATAATCAGAAGTTCAGAGGCAG

GGCCCAACTGACTGTAGACACATCCTCCAGCACAGCCTACATGCAGTTCAGCAGCCTGACAACT

GATGACTCTGCCATCTATTACTGTGCAAGACACGTGAGGGGTTACCATCCTATGGACTACTGGG

GTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACGAAGGGCCCATCCGTCTTCCCCCTGGC

GCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTC

CCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGG

CTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTG

GGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGA

GTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGGGACCAT

CAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCAC

GTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGG

CGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGT

GGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGT

CTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCG

AGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCT

GACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCA

GCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTAC

AGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATG

CATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGCTAGCT

GA

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

WIAWIYAGTGGITYNQKFRGRAQLTVDTSSSTAYMQFSSLTTDDSAIYYCARHVRGYHPMDYWGQ

GTSVTVSSAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS

GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPK

DTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDW

LNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE

WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

KAS

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

TGTGATGACCCAGTCTCAAAAATTCATGTCCGCATCAGTAGGGGACAGGGTCAGCGTCACCTGC

AGGGCCAGTCAGAATGTGGTTACTAATGTAGGCTGGTATCAACAGAAACCAGGGCAATCTCCT

AAAGTACTGATTTACTCGGCATCCTTCCGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTG

GATCTGGGACAGATTTCACTCTCACCATCACCAATGTGCAGTCTGAAGACTTGGCAGAGTATTT

CTGTCAGCAATATAACAACTATCCGTACACGTTCGGAGGGGGGACCAAGCTCGAGATCAAACG

AACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTG

CCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGA

TAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCAC

CTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC

CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG

TTAG

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

KVLIYSASFRYSGVPDRFTGSGSGTDFTLTITNVQSEDLAEYFCQQYNNYPYTFGGGTKLEIKRTVAA

PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL

TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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

AACTGCAGCAGCCTGGGGCTGAACTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGG

CTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGAGAAGGCCT

TGAGTGGATTGGAGAGATTAATCCTAGCTACGGTCGTACTGACTACAATGGGAAGTTCAAGAAC

AAGGCCACACTGACTGTAGCCAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACAT

CTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGAGATTACTACGGTAGTAGCTCGTTTGCTTA

CTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCAAAACAAAGGGCCCATCCGTCTTCCCC

CTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACT

ACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTT

CCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC

AGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGAC

AAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCGAAGGGG

GACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGA

GGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGT

GGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTA

CCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGC

AAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAG

CCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTC

AGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT

GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC

TCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCG

TGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAAGC

TAGCTGA

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

WIGEINPSYGRTDYNGKFKNKATLTVAKSSSTAYMQLSSLTSEDSAVYYCARGDYYGSSSFAYWGQ

GTLVTVSAAKTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS

SGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKP

KDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQD

WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAV

EWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL

GKAS

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

TGTGATGACCCAGTCTCAAAAATTCATGTCCACATCATTAGGAGACAGGGTCAGCGTCACCTGC

AAGGCCAGTCAGAATGTGGGTACTAATGTAGCCTGGTATCAACAGAAACCAGGGCACTCTCCT

AAAGCACTGATTTACTCGGCATCCTACCGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTG

GATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTTTTT

CTGTCAGCAATATAACAACTATCCGTACACGTTCGGAGGGGGGACCACGCTCGAGATCAAACG

AACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTG

CCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGA

TAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCAC

CTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTATGC

CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG

TTAG

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

KALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEFFCQQYNNYPYTFGGGTTLEIKRTVAA

PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL

TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

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)
DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSG

TDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN

NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS

SPVTKSFNRGEC

hAnti-CD40VH3-LV-hIgG4H-C-Flex-v1-HelB-f1-ProtB-f2-NS5BPalm
(SEQ ID NO: 159)
EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVK

GRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSAKTKGPSVFPL

APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTK

TYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS

QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI

EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD

SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

ASVTVPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVAL

GINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTLP

QDAVSRTQRRGRTGRGKPGIYRFVAPGERAS ASTPCTCGSSDL

YLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGHAVGIFRAAVCTRGVAKAVD

FIPVENLETTMRSPVFTDNSSPPAVPQSAS ASVLDSHYQDVLKE

VKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPGQRVEFLVQAWKSKKTPMGFSYDTRCFDSTVTESDI

RTEEAIYQCCDLDPQARVAIKSLTERLYVGRCRASGVLTTSCGNTLTCYIKARAACRAAGLQDCTMLVCGD

DLVVICESAGVQEDAASLRAFTEAMTRYSAPPGDPPQPEYDLELITAS

Humanized anti-DCIR-HCV 1^stgeneration 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)
DIVMTQSPDSLAVSLGERATINCRASESIHSYGNSFLHWYQQKPGQPPKLLIYLASNLESGVPSRFSG

SGSRTDFTLTISSLQPEDFATYYCQQNNEDPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV

VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT

HQGLSSPVTKSFNRGEC

hAnti-DCIRVH1-LV-hIgG4H-C-Flex-v1-HelB-f1-ProtB-f2-NS5BPalm
(SEQ ID NO: 161)
QVTLKESGPAIVKPTQTLTLTCSFSGFSLSTSGMGLSWIRQPSGKALEWLAHIYWDDDKRYNPSLKS

RLTISKDTSKNQVVLTMTIVDTVDAATYYCARSSHYYGYGYGGYFDVWGQGTTVTVSSAKTKGPS

VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS

LGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVV

VDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG

LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP

PVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

ASVTVPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKL

VALGINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIET

TTLPQDAVSRTQRRGRTGRGKPGIYRFVAPGERAS ASTPCTCG

SSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGHAVGIFRAAVCTRGVA

KAVDFIPVENLETTMRSPVFTDNSSPPAVPQSAS ASVLDSHYQ

DVLKEVKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPGQRVEFLVQAWKSKKTPMGFSYDTRCFDST

VTESDIRTEEAIYQCCDLDPQARVAIKSLTERLYVGRCRASGVLTTSCGNTLTCYIKARAACRAAGLQDCTM

LVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPPGDPPQPEYDLELITAS

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)
DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSG

TDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN

NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS

SPVTKSFNRGECASVTVPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELAAKLVA

LGINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTFTIETTTL

PQDAVSRTQRRGRTGRGKPGIYRFVAPGERAS

hAnti-CD40VH3-LV-hIgG4H-C-Flex-v1-ProtB-f1-NS5BPalm
(SEQ ID NO: 163)
EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVK

GRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSAKTKGPSVFPL

APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTK

TYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS

QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI

EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD

SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

ASTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPA

GHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQSAS

ASVLDSHYQDVLKEVKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPGQRVEFLVQAW

KSKKTPMGFSYDTRCFDSTVTESDIRTEEAIYQCCDLDPQARVAIKSLTERLYVGRCRASGVLTTSCGNTLT

CYIKARAACRAAGLQDCTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPPGDPPQPEYDLELITA

S

Humanized anti-DCIR-HCV 2^ndgeneration vaccine is: [hAnti-DCIRVK4-LV-hIgGK-C-ViralHCVhelicasefgtB]×[hAnti-D CIRVH1-LV-hIgG4H-C—F lex-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)
DIVMTQSPDSLAVSLGERATINCRASESIHSYGNSFLHWYQQKPGQPPKLLIYLASNLESGVPSRFSG

SGSRTDFTLTISSLQPEDFATYYCQQNNEDPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV

VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT

HQGLSSPVTKSFNRGECASVTVPHPNIEEVALSTTGEIPFYGKAIPLEVIKGGRHLIFCHSKKKCDELA

AKLVALGINAVAYYRGLDVSVIPTSGVVVVVATDALMTGFTGDFDSVIDCNTCVTQTVDFSLDPTF

TIETTTLPQDAVSRTQRRGRTGRGKPGIYRFVAPGERAS

hAnti-DCIRVH1-LV-hIgG4H-C-Flex-v1-ProtB-f1-NS5BPalm
(SEQ ID NO: 165)
QVTLKESGPAIVKPTQTLTLTCSFSGFSLSTSGMGLSWIRQPSGKALEWLAHIYWDDDKRYNPSLKS

RLTISKDTSKNQVVLTMTIVDTVDAATYYCARSSHYYGYGYGGYFDVWGQGTTVTVSSAKTKGPS

VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS

LGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVV

VDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG

LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP

PVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKAS

ASTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLL

CPAGHAVGIFRAAVCTRGVAKAVDFIPVENLETTMRSPVFTDNSSPPAVPQSAS

ASVLDSHYQDVLKEVKAAASKVKANALYDVVSKLPLAVMGSSYGFQYSPGQRVEFLV

QAWKSKKTPMGFSYDTRCFDSTVTESDIRTEEAIYQCCDLDPQARVAIKSLTERLYVGRCRASGVLTTSCGN

TLTCYIKARAACRAAGLQDCTMLVCGDDLVVICESAGVQEDAASLRAFTEAMTRYSAPPGDPPQPEYDLEL

ITAS

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. 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.

2. The composition of claim 1, further comprising at least one Toll-Like Receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists.

3. The composition of claim 1, further comprising 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.

4. The composition of claim 1, wherein the antibody or fragment specific for the DC is specific for a DC specific cell surface receptor 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.

5. The composition of claim 1, wherein the HCV antigens comprise a peptide sequence derived from a HCV 1a genotype protein or a fragment thereof.

6. The composition of claim 1, wherein 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.

7. The composition of claim 1, wherein 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, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, or a fragment thereof.

8. The composition of claim 1, wherein the composition comprises a recombinant antibody that comprises a fusion protein and the one or more HCV antigens at a C-terminal position relative to the one or more antibody or fragment thereof; a recombinant antibody or fragment thereof specific for the DC and one or more HCV antigens are fused to a C-terminus of a heavy chain of the antibody; a recombinant antibody or fragment thereof specific for the DC and one or more HCV antigens are fused to a C-terminus of a light chain of the antibody; or a recombinant antibody or fragment thereof specific for the DC and three HCV antigens or antigenic domains fused to the C-terminus of the heavy chain of the antibody or two HCV antigens or antigenic domains fused to the heavy chain of the antibody and one HCV antigen or antigenic domain fused to the light chain of the antibody.

9. The composition of claim 1, wherein 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-linkerB-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.

10. The composition of claim 1, wherein 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; or 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.

11. The composition of claim 1, wherein 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.

12. The composition of claim 1, wherein the DC-specific antibody is humanized.

13. The composition of claim 1, wherein the one or more HCV antigens are selected from antibodies having a heavy and a light chain combination of sequences SEQ ID NOS: 80 and 81; 82 and 83; 84 and 85; 86 and 87; 88 and 89; 90 and 91; 92 and 93; 94 and 95; 96 and 97; 98 and 99; 100 and 101; 102 and 103; 104 and 105; 106 and 107; 108 and 109; 110 and 111; 112 and 113; 114 and 115; 116 and 117; 118 and 119; 120 and 121; 122 and 123; 124 and 125; 126 and 127; 128 and 129; 130 and 131; 132 and 134; 136 and 137; 138 and 139; 140 and 141; 158 and 159; 160 and 161; 162 and 163; 164 and 165.

14. A vaccine comprising:

one or more HCV antigens attached to the one or more antibodies or fragments thereof and optionally a 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 a subject in need of immunostimulation.

15. The vaccine of claim 14, further comprising at least one Toll-Like Receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, and TLR8 agonists.

16. The vaccine of claim 14, wherein the HCV antigen comprises a peptide sequence derived from a HCV 1a genotype protein or a fragment thereof, protein E1, envelope protein E2, non-structural protein NS3, non-structural protein NS4b, non-structural protein NS5b, and a fragment thereof.

17. The vaccine of claim 14, wherein the one or more HCV antigens 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, or a fragment thereof.

18. The vaccine of claim 14, wherein the composition comprises a recombinant antibody that comprises a fusion protein and the one or more HCV antigens are at a C-terminal position relative to the one or more antibody or fragment thereof within a fusion protein; one or more HCV antigens are fused to a C-terminus of a heavy chain of the antibody; 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 dendritic cell; or one or more HCV antigens or antigenic domains fused to a C-terminus of the heavy chain of the antibody and at least one HCV antigen or antigenic domain fused to the C-terminus of the light chain of the antibody.

19. The vaccine of claim 14, wherein the vaccine comprises a recombinant antibody or fragment thereof specific for the DC and three HCV antigens or antigenic domains fused to the C-terminus of the heavy chain of the antibody or two HCV antigens or antigenic domains fused to the heavy chain of the antibody and one HCV antigen or antigenic domain fused to the light chain of the antibody.

20. The vaccine of claim 14, wherein 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-linkerB-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.

21. The vaccine of claim 14, wherein the one or more HCV antigens 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.

22. The vaccine of claim 14, wherein the DC-specific antibody is humanized.

23. The vaccine of claim 14, wherein the vaccine comprises an antibody with a heavy and a light chain combination of sequences SEQ ID NOS: 80 and 81; 82 and 83; 84 and 85; 86 and 87; 88 and 89; 90 and 91; 92 and 93; 94 and 95; 96 and 97; 98 and 99; 100 and 101; 102 and 103; 104 and 105; 106 and 107; 108 and 109; 110 and 111; 112 and 113; 114 and 115; 116 and 117; 118 and 119; 120 and 121; 122 and 123; 124 and 125; 126 and 127; 128 and 129; 130 and 131; 132 and 134; 136 and 137; 138 and 139; 140 and 141; 158 and 159; 160 and 161; 162 and 163; 164 and 165.

24. A Hepatitis C vaccine (HCV) comprising a fusion protein comprising:

one or more antibodies or fragments thereof specific for a dendritic cell (DC);

one or more HCV antigens located C-terminal of the antibodies or fragments thereof;

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 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.

25. The vaccine of claim 24, wherein 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.

26. The vaccine of claim 24, wherein the antibody or fragment specific for the DC 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.

27. A method for increasing effectiveness of Hepatitis C virus (HCV) antigen presentation by an antigen presenting cell (APC) comprising the steps of:

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;

providing one or more APCs; and

contacting the APC with the conjugate, wherein the antibody-antigen complex is processed and presented for T cell recognition.

28. The method of claim 27, wherein the antigen presenting cell comprises a dendritic cell (DC).

29. The method of claim 27, 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, CD 14, CD 15, CD 16, 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.

30. The method of claim 27, 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: 6, 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.

31. The method of claim 27, wherein the DC-specific antibody is humanized.

32. A method for increasing effectiveness of antigen presentation by an antigen presenting cell (APC) comprising the steps of:

isolating and purifying one or more dendritic cell (DC)-specific antibodies or a fragments thereof;

providing one or more HCV antigens or antigenic peptides;

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

contacting the antigen presenting cell with the conjugate, wherein the antibody-antigen complex is processed and presented for T cell recognition.

33. The method of claim 32, further comprising the optional steps 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 antibody-antigen conjugate and the TLR agonist prior to contacting the antigen presenting cells; and

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.

34. The method of claim 32, wherein the APC comprises a dendritic cell (DC).

35. The method of claim 32, 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, CD 15, 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.

36. The method of claim 32, 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.

37. The method of claim 32, wherein the one or more HCV antigens 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 a fragment thereof.

38. The method of claim 32, wherein the DC-specific antibody is humanized.

39. The method of claim 32, further comprising the optional 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.

40. The method of claim 32, further comprising 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.

41. A Hepatitis C virus antigen presenting dendritic cell (DC) comprising 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.

42. 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;

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.

43. The method of claim 42, further comprising administering to the subject an effective amount of IFNA, Ribavirin, or a combination thereof.

44. 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:

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.

45. The vaccine of claim 44, wherein w comprises the HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, Palm, E1b, and E2.

46. The vaccine of claim 44, wherein x comprises the HCV antigenic domains selected from the group consisting of HelC, HelA, Palm, ProtA, ProtB, and E1b.

47. The vaccine of claim 44, wherein y comprises the HCV antigenic domains selected from the group consisting of Palm, ProtB, and Protb.

48. The vaccine of claim 44, wherein z comprises HCV antigenic domains selected from E2, ProtA, and HelB.

49. The vaccine of claim 44, wherein the one or more HCV antigens or antigenic domains are linked or attached to one another by one or more flexible linkers.

50. The vaccine of claim 44, wherein the vaccine comprises H-ProtA, H-ProtB, H-HelB, H-Palm, H-E1b, H-E2, H-HelB-HelC, H-HelB-HelA, H-HelB-Palm, H-Palm-HelB, H-HelB-ProtB, H-ProtB-HelB, H-ProtA-ProtB, H-ProtB-ProtA, H-ProtB-Palm, H-Palm-ProtB, H-ProtA-HelB, H-HelB-ProtA, H-E2-E1b, H-HelB-ProtB-Palm, H-HelB-Palm-ProtB, H-ProtB-HelB-Palm, H-ProtB-Palm-HelB, H-Palm-HelB-ProtB, H-Palm-ProtB-HelB, H-HelB-ProtB-Palm-E2, or any combinations thereof.

51. 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.

52. The composition of claim 51, wherein w, x, y, and z comprise HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, HelA, HelC, Palm, E1a, E1b, and E2.

53. The composition of claim 51, wherein the vaccine comprises L-ProtA-HelA-E1b-HelB.

54. 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.

55. The composition of claim 54, wherein w, x, y, and z comprise HCV antigenic domains selected from the group consisting of ProtA, ProtB, HelB, HelA, HelC, Palm, E1a, E1b, and E2.

56. The composition of claim 54, wherein the vaccine comprises

57. The composition of claim 54, wherein the vaccine comprises