DNA antibody and immunogen constructs for use in methods of treating and preventing influenza

Abstract

Disclosed herein are compositions comprising a recombinant nucleic acid sequence for generating one or more synthetic antibodies, including a DNA-encoded monoclonal antibody targeting influenza A virus hemagglutinin (HA) protein and a DNA- encoded monoclonal antibody targeting influenza A virus neuraminidase (NA) protein, compositions comprising a recombinant nucleic acid sequence for generating one or more immunogen, including influence A nucleoprotein (NP) and influenza A virus hemagglutinin (HA) protein. The disclosure also provides a method of preventing and / or treating influenza in a subject using said composition.

Description

[0001] DNA ANTIBODY AND IMMUNOGEN CONSTRUCTS FOR USE IN

[0002] METHODS OF TREATING AND PREVENTING INFLUENZA

[0003] CROSS REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 637,195, filed April 22, 2024, U.S. Provisional Application No. 63 / 644,230, filed May 08, 2024, U.S. Provisional Application No. 63 / 710,420, filed October 22, 2024, and U.S. Provisional Application No. 63 / 717,009, filed November 06, 2024, each of which is hereby incorporated by reference herein in its entirety.

[0005] BACKGROUND

[0006] Despite promising innovations, influenza vaccines and antiviral drugs do not provide full protection from seasonal infection, and provide little immediate defense against novel and potentially pandemic viral strains. Broadly cross-protective monoclonal antibodies have been developed with the aim of providing protection against highly divergent influenza viruses.

[0007] Thus, there is a need in the art for improved compositions and methods for the treatment of influenza.

[0008] SUMMARY

[0009] In some embodiments, the invention provides an anti-HA (influenza a virus hemagglutinin) antibody or fragment thereof.

[0010] In some embodiments, the anti-HA antibody comprises a light chain selected from the group consisting of: a) SEQ ID NO:26, SEQ ID NO:28. SEQ ID NO:30, SEQ ID NO: 32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, and SEQ ID NO:46; b) an amino acid sequence at least 90% identical to SEQ ID NO: 26, SEQ ID NO:28, SEQ ID NO:30. SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36. SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, and SEQ ID NO:46; c) an amino acid sequence at least 70% of the length of SEQ ID NO:26. SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32. SEQ ID NO:34, SEQ ID NO:36. SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, and SEQ ID NO:46; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34. SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44. and SEQ ID NO:46.

[0011] In some embodiments, the anti-HA antibody comprises a heavy chain selected from the group consisting of: a) SEQ ID NO:2, SEQ ID NO: 4. SEQ ID NO:6. SEQ ID NO:8. SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO:20, SEQ ID NO:22, and SEQ ID NO:24; b) an amino acid sequence at least 90% identical to SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:8. SEQ ID NO:10, SEQ ID NO: 12, SEQ ID NO: 14. SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO:20. SEQ ID NO:22, and SEQ ID NO:24; c) an amino acid sequence at least 70% of the length of SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:8. SEQ ID NO:10, SEQ ID NO: 12, SEQ ID NO: 14. SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO 20, SEQ ID NO:22, and SEQ ID NO:24; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14. SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO:20, SEQ ID NO:22. and SEQ ID NO:24.

[0012] In some embodiments, the antibody is selected from the group consisting of a humanized antibody, a chimeric antibody, a fully human antibody, and an antibody mimetic.

[0013] In some embodiments, the invention relate to a nucleic acid molecule encoding an anti-HA antibody.

[0014] In some embodiments, the nucleotide sequence encoding the light chain of an anti-HA antibody is selected from the group consisting of: a) SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31,

[0015] SEQ ID NO 33, SEQ ID NO:35. SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, and SEQ ID NO:45; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29. SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35. SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, and SEQ ID NO:45; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:25. SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO 31, SEQ ID NO:33, SEQ ID NO:35. SEQ ID NO:37, SEQ ID NO:39. SEQ ID NO:41, SEQ ID NO:43, and SEQ ID NO:45; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO 39, SEQ ID NO:41, SEQ ID NO:43, and SEQ ID NO:45.

[0016] In some embodiments, the nucleotide sequence encoding the heavy chain of an anti-HA antibody is selected from the group consisting of: a) SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NON. SEQ ID NO:11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO:17, SEQ ID NO: 19, SEQ ID NO:21, and SEQ ID NO:23; b) a nucleic acid sequence at least 90% identical to SEQ ID NO: 1 , SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO 19, SEQ ID NO:21, and SEQ ID NO:23: c) a nucleic acid sequence at least 70% of the length of SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO 19, SEQ ID NO:21, and SEQ ID NO:23; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NON, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21. and SEQ ID NO:23.

[0017] In some embodiments, the invention relates to an anti-NA (influenza a vims neuraminidase) antibody or fragment thereof.

[0018] In some embodiments, the anti-NA antibody comprises a light chain selected from the group consisting of: a) SEQ ID NO:49 and SEQ ID NO:53; b) an amino acid sequence at least 90% identical to SEQ ID NO: 49 and

[0019] SEQ ID NO:53; c) an amino acid sequence at least 70% of the length of SEQ ID NO:49 and SEQ ID NO:53; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 49 and SEQ ID NO: 53.

[0020] In some embodiments, the anti-NA antibody, or fragment thereof, comprises a heavy chain selected from the group consisting of: a) SEQ ID NO: 47 and SEQ ID NO:51; b) an amino acid sequence at least 90% identical to SEQ ID NO:47 and SEQ ID NO:51; c) an amino acid sequence at least 70% of the length of SEQ ID NO:47 and SEQ ID NO:51; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 47 and SEQ ID NO:51.

[0021] In some embodiments, the anti-NA antibody or fragment thereof is selected from the group consisting of a humanized antibody, a chimeric antibody, a fully human antibody, and an antibody mimetic.

[0022] In some embodiments, the invention provides a nucleic acid molecule encoding an anti-NA antibody, or fragment thereof.

[0023] In some embodiments, the nucleotide sequence encoding the light chain of an anti-NA antibody is selected from the group consisting of: a) SEQ ID NO:50 and SEQ ID NO:54; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:50 and SEQ ID NO:54; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:50 and SEQ ID NO:54; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:50 and SEQ ID NO:54.

[0024] In some embodiments, the nucleotide sequence encoding the heavy chain of an anti-NA antibody is selected from the group consisting of: a) SEQ ID NO:48 and SEQ ID NO:52; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:48 and

[0025] SEQ ID NO:52; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:48 and SEQ ID NO:52; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:48 and SEQ ID NO:52.

[0026] In some embodiments, the invention provides an influenza A nucleoprotein (NP) polypeptide.

[0027] In some embodiments, the NP polypeptide comprises an amino acid sequence selected from the group consisting of a) SEQ ID NO: 56 or SEQ ID NO: 58; b) an amino acid sequence at least 90% identical to SEQ ID NO: 56 or SEQ ID NO: 58; c) an amino acid sequence at least 70% of the length of SEQ ID NO:

[0028] 56 or SEQ ID NO: 58; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 56 or SEQ ID NO: 58.

[0029] In some embodiments, the invention provides a nucleic acid molecule encoding an NP polypeptide.

[0030] In some embodiments, the nucleotide sequence is selected from the group consisting of: a) SEQ ID NO:55 or SEQ ID NO:57; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:55 or SEQ ID NO:57; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:55 or SEQ ID NO:57; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:55 or SEQ ID NO:57.

[0031] In some embodiments, the invention provides an influenza A hemagglutinin (HA) protein polypeptide.

[0032] In some embodiments, the polypeptide comprises an amino acid sequence selected from the group consisting of: a) SEQ ID NO: 60; b) an amino acid sequence at least 90% identical to SEQ ID NO: 60; c) an amino acid sequence at least 70% of the length of SEQ ID NO:

[0033] 60; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 60.

[0034] In some embodiments, the invention provides a nucleic acid molecule encoding an HA polypeptide.

[0035] In some embodiments, the nucleotide sequence is selected from the group consisting of: a) SEQ ID NO:59; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:59; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:59; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:59.

[0036] In some embodiments, any of the the nucleotide sequences described herein encodes a leader sequence.

[0037] In some embodiments, any of the nucleic acid molecules described herein comprise an expression vector.

[0038] In some embodiments, the invention provides a composition comprising any nucleic acid molecule described herein.

[0039] In some embodiments, the invention relates to a composition comprising: a) at least one anti-HA antibody or fragment thereof: b) at least one anti-NA antibody or fragment thereof; c) at least one NP polypeptide; or d) at least one HA polypeptide.

[0040] In some embodiments, the composition further comprises a pharmaceutically acceptable excipient.

[0041] In some embodiments, the invention provides a method of preventing or treating a disease in a subject, the method comprising administering to the subject the antibody or antibody fragment of the invention, the polypeptide of the invention, the nucleic acid molecule of the invention, a composition of the invention, or any combination thereof.

[0042] In some embodiments, the disease is influenza. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] The following description of exemplary embodiments of the invention will be better understood when read in conjunction with the appended drawings. It should be understood, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

[0044] Figure 1 depicts representative exemplary data demonstrating that dMAb- FISW086 has a comparable neutralization IC50 titre to dMAb-2-12C. Neutralization was performed on heat inactivated sera incubated with 100 TCID50 of H IN 1 / Bethesda (received from NIH). An overnight assay was performed according to the WHO influenza assay guidelines. Presence or absence of infection was detected by an anti-influenza NP ELISA.

[0045] Figure 2 depicts representative exemplary data evaluating dMAb-FISW086 in a DBA / 2 / CalO9 lethal model. These data demonstrated that dMAb-FISW086 is protective and prevents weight loss at very low doses.

[0046] Figure 3 depicts representative exemplary data evaluating FISW086 dMAb dosage. No treatment: interstitial pneumonia with dense cellular infiltrate, stains positive for IAV-NP. dMAb-FISW086 12.5 pg and 25 pg: mild interstitial pneumonia and stain negative for IAV-NP. dMAb-FISW086 6.25 pg and 3. 12: mild interstitial pneumonia and stains positive for IAV-NP around airways. Lung analysis enables a better visualization of dMAb effectiveness to support down-selection and additional development.

[0047] Figure 4 depicts representative exemplar}- data evaluating dMAb-LPAF-021 in a DBA / 2 / CalO9 lethal model. These data demonstrate that dMAb-LPAF021 shows protection against weight loss and signs of disease at very low doses. LPAF-021 is an additional mAb / dMAb that binds to a different epitope compared to FISW086.

[0048] Figure 5 depicts exemplary data evaluating LPAF021 dMAb dosage. No treatment: interstitial pneumonia with dense cellular infiltrate, stains positive for IAV-NP. dMAb- LPAF021 25 pg: mild interstitial pneumonia and stains negative for IAV-NP. dMAb- LPAF021 12.5, 6.25 pg and 3. 12: moderate to severe interstitial pneumonia and stains positive for IAV-NP. Lung analysis enables a better visualization of dMAb effectiveness to support down-selection and additional development.

[0049] Figure 6 depicts representative exemplary data evaluating dMAb cocktail delivery in the DBA / 2 lethal model. These data demonstrate that that a combination of the two dMAbs may improve protection synergistically at lower doses. LPAF-021 is an additional mAb / dMAb that binds to a different epitope compared to FISW086. Figure 7 depicts exemplary data evaluating combination dMAb dosage. No treatment: interstitial pneumonia with dense cellular infiltrate, stains positive for IAV-NP. A combination of the 2 dMAbs protects against pneumonia at 25+25 pg and 12.5+12.5 pg and stain negative for NP. The 6.25+6.25 group shows mild pneumonia and stains positive for NP in small areas of the lung. The 3.12 + 3.12 group shows mild pneumonia in small areas of the lung and stains negative for flu NP.

[0050] Figure 8 depicts representative exemplary data demonstrating that dMAb- AF9C and dMAb-AG7C protect against lethal challenge and that dMAb-AG7C affords the better protection in the lungs.

[0051] Figure 9, comprising Figure 9A-Figure 9H, depicts the design and in vitro expression of VACC-NPXimmunogens. Figure 9A and 9D depict NP amino acid alignments of seasonal A / H1N1 and A / H3N2 vaccine strains (GISAID.org). Figure 9B and 9E depict unrooted phylogenetic trees for A / H1N1 and A / H3N3 vaccine strains. Figure 9C and 9F depict plasmid maps of pVACC-NPH1and pVACC-NPH3synthetic DNA constructs. Figure 9G depicts the results of a western blot of pVACC-NPH1and pVACC-NPH3HEK29T supernatants probed for anti-IAV-NP. Figure 9H depicts Immunofluorescence staining of HEK293T cells transfected with pVACC-NPxplasmids and stained for IAV-NP.

[0052] Figure 10, comprising Figure lOA-Figure 10H, depicts the results of example experiments demonstrating a single immunization with VACC-NPXconstructs is immunogenic and protects against mortality in an (H1N1) pdm09 mouse infection model. Figure 10A depicts the experimental design, wherein C57BL / 6 mice were immunized with lOpg of pVACC-NPxplasmids and sacrificed fourteen days post-immunization for cellular analyses. Figure 10B and IOC depict IFNy spot-forming units (SFUs) in spleens following stimulation with H1NP peptides or H3NP peptides (n=5 mice per group). Figure 10D-10H depicts results of example experiments wherein DBA / 2 mice received a single administration of the pVACC-NPH1or pVACC-NPH3synthetic DNA vaccines (lOpg, n=10 mice / group). After 14 days, the mice were intranasally challenged with 10LD50 of H1N1 A / Califomia / 07 / 2009 X179A and monitored daily until day 21 post-challenge. On day 6 post-infection, a subset of mice (n=3) was euthanized lungs were collected and processed for histopathological analyses. Figure 10D depicts the experimental design, Figure 10E depicts survival curves, and Figure 10F depicts weight loss. Figure 10G depicts gematoxylin and eosin staining, and Figure 10H depicts IAV-NP immunohistochemistry staining of lung sections from representative mice at 6 days post-infection. Scale bars equal 2.5 mm on whole slide lung images **p<0.0L ***p<0.001, ****p<0.0001 by Two-way ANOVA.

[0053] Figure 11, comprising Figure 1 lA-Figure 11C, depicts the results of example experiments demonstrating pVACC-NPximmunogens are amenable to co-delivery with HA immunogens. Figure 11A depicts the experimental design wherein C57BL / 6 mice were immunized with 2 pg plasmid-encoded California 2008 HA (pHAH1) and lOpg of pVACC- NPH3separately, together, or co-immunized with pHAH1, pVACC-NPH3, and 0.5pg of IL-12 (combo), and sacrificed fourteen days post-immunization for cellular analyses. Figure 11B depicts INFy+effector CD8+ T cells in spleens following stimulation with H3 NP peptides, INFy CD107+effector CD8+ T cells in spleens following stimulation with H3 NP peptides, INFy+effector CD8+ T cells in lungs following stimulation with H3 NP peptides and INFy+CD107+effector CD8+ T cells in lungs following stimulation with H3 NP peptides. Figure 11C depicts INFy effector CD8+ T cells in spleens following stimulation with H1N1 HA peptides, INFy+CD107+effector CD8+ T cells in spleens following stimulation with H1N1 HA peptides. INFy+effector CD8+ T cells in lungs following stimulation with H IN 1 HA peptides, and INFy+CD107+effector CD8+ T cells in lungs following stimulation with H1N1 HA peptides. Data are representative of one independent experiment with N=5 / group. Bars represent the group mean; error bars represent SEM or SD . *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 by two-way ANOVA or Kruskal -Wallis ANOVA.

[0054] Figure 12, comprising Figure 12A-Figurel2E, depicts the results of example experiments demonstrating HA and NP combination improves protection from IAV induced morbidity. Figure 12A depicts the experimental overview. DBA / 2 mice were immunized one with lOpg of pHAH1alone. 1 Opg pVACC-NPH3alone, or co-immunized with pHAH1, pVACC-NPH3and 0.5pg of IL-12 (combo), and challenged with 100LD50 A / Califomia / 2009 IAV virus 14 days later. Lungs were collected from 3 representative animals per group 6 days post-challenge and the remaining animals were monitored daily for weight loss and survival. Figure 12B depicts body weight as a percent of starting weight. Figure 12C depicts survival probability. Figure 12D depicts H&E stained representative lungs from animals euthanized at day 6 post-challenge. Figure 12E depicts NP antigen-stained representative lungs from animals euthanized at day 6 post-challenge. Data are representative of one independent experiment with N=10 / group. Bars represent the group mean; error bars represent SEM.

[0055] Figure 13, comprising Figure 13A-Figure 13G, depict the results ofRNA-seq analysis of H1HA vs. H3HP DNA vaccines during infection. Figure 13A depicts the experimental design wherein Mice (DBA / 2) were immunized with one dose of pHAH1or pVACC-NPH3and 14 days later, challenged with 10 LD50 of A / Cahfomia / 2009 IAV virus. Lungs were harvested at day-6 p.i. and processed for bulk RNA-seq. Figure 13B depicts PCA analysis showing an overlapping cluster of the two vaccinated groups distinct from the nonimmunized infected mice. Figure 13C depicts gene analysis showing all genes expressed in the immunized and infected groups. Figure 13D and 13E depict volcano plots for mice immunized with pHAH1or pVACC-NPH3and show the total number of up and down regulated genes. Figure 13F and 13G depict ingenuity pathway analysis showing activated and inhibited pathways related to viral infection, immune cells, cytokines, and others.

[0056] Figure 14, comprising Figure 14A-Figure 14J depict the results of example experiments demonstrating VACC-NP immunogens induce robust and durable cellular responses in vivo. Figure 14A depicts the experimental design, wherein female C57BL / 6 mice were immunized once with 10pg of vaccine-aligned consensus DNA plasmid constructs (pVACC-NPHx) representing nucleoproteins (NP) from HlNl (pVACC-NPH1) or H3N2 (pVACC-NPH3) alone, or co-immunized with pVACC-NPHxand 0.5pg of plasmid-encoded IL-12 (+pTL- 12). Figure 14B depicts Hl NP-specific IFNy spot-forming units (SFU) in spleens. Figure 14C depicts Hl NP-specific IFNy SFU in lungs. Figure 14D depicts H3 NP- specific IFNy SFU in spleens. Figure 14E depicts H3 NP-specific IFNy SFU in lungs. Figure 14F depicts the experimental design, wherein Mice were immunized twice as in Figure 14A. separated by three weeks and cellular responses were evaluated seven days post-final immunization. Figure 14G depicts Hl NP-specific IFNy spot-forming units (SFU) in spleens. Figure 14H depicts Hl NP-specific IFNy SFU in lungs. Figure 141 depicts H3 NP-specific IFNy SFU in spleens. Figure 14J depicts H3 NP-specific IFNy SFU in lungs. Data are representative of two independent experiments with N=5 / group. Bars represent the mean; error bars represent SEM or SD. **p<0.01, ***p<0.001, ****p<0.0001 by Two-way ANOVA.

[0057] Figure 15 depicts a summary of strains used in analysis.

[0058] Figure 16 depicts multiple sequence alignment of NP proteins of seasonal A / H1N1 vaccine strains, performed in Geneious Prime. Dots represent complete identity of the residue in all the sequences. The green bar on top represents the level of conservation of the residues between strains, deep green indicates most conserved, yellow indicates a poorly- conserved position. Figure 17 depicts multiple sequence alignment of NP proteins of seasonal A / H3N2 vaccine strains, performed in Geneious Prime. Dots represent complete identity of the residue in all the sequences. The green bar on top represents the level of conservation of the residues between strains, deep green indicates most conserved, yellow indicates a poorly conserved position.

[0059] Figure 18, comprising Figure 18A-Figure 18E, depicts the results of example experiments demonstrating the identification of pVACC-NPximmunodominant T cell epitopes. Figure 18A depicts the experimental design, wherein C57BL / 6 mice were immunized twice, separated by three weeks, with I Opg of pVACC-NPx. Matrix peptide pools were used to stimulate isolated splenocytes. Figure 18B depicts H1N1-NP specific IFNy secretion as measured by ELISpot. Figure 18C depicts identified H INI -NP immunodominant peptides. Figure 18D depicts H3N2-NP specific IFNy secretion as measured by ELISpot. Figure 18E depicts identified H3N2-NP immunodominant peptides.

[0060] Figure 19, comprising Figure 19A-Figure 19C, depicts the results of example experiments demonstrating pHAHl is sub-protective in high-dose I AV challenge. Figure 19A depicts the experimental design, wherein mice were immunized once with lOpg, Ipg, or 0.5pg of plasmid-encoded A / Califomia / 2009 HA (pHAHl) and challenged with 100LD50 of matched virus fourteen days later. Figure 19B depicts the surv ival probability. Figure 19C depicts body weights post-challenge as percent of starting weight.

[0061] DETAILED DESCRIPTION

[0062] The invention relates to compositions comprising a recombinant nucleic acid sequence encoding an antibody, a fragment thereof, a variant thereof, or a combination thereof. In some embodiments, the antibody is an anti-HA antibody. In some embodiments, the antibody is an anti-NA antibody The composition can be administered to a subject in need thereof to facilitate in vivo expression and formation of a synthetic antibody.

[0063] In particular, the heavy chain and light chain polypeptides expressed from the recombinant nucleic acid sequences can assemble into the synthetic antibody. The heavychain polypeptide and the light chain polypeptide can interact with one another such that assembly results in the synthetic antibody being capable of binding the antigen, being more immunogenic as compared to an antibody not assembled as described herein, and being capable of eliciting or inducing an immune response against the antigen. Additionally, these synthetic antibodies are generated more rapidly in the subject than antibodies that are produced in response to antigen induced immune response. The synthetic antibodies are able to effectively bind and neutralize a range of antigens. The synthetic antibodies are also able to effectively protect against and / or promote survival of disease.

[0064] Definitions

[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0066] The terms “comprise(s),” “include(s),” ‘“having;’ “‘has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “and” and “the” include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments “comprising.” “consisting of’ and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not.

[0067] “Antibody” may mean an antibody of classes IgG, IgM, IgA, IgD or IgE, or fragments, fragments or derivatives thereof, including Fab, F(ab')2, Fd, and single chain antibodies, and derivatives thereof. The antibody may be an antibody isolated from the serum sample of mammal, a polyclonal antibody, affinity purified antibody, or mixtures thereof which exhibits sufficient binding specificity to a desired epitope or a sequence derived therefrom.

[0068] “Antibody fragment” or “fragment of an antibody” as used interchangeably herein refers to a portion of an intact antibody comprising the antigen-binding site or variable region. Examples of antibody fragments include, but are not limited to. Fab fragments. Fab' fragments, Fab'-SH fragments, F(ab')2 fragments, Fd fragments, Fv fragments, diabodies, single-chain Fv (scFv) molecules, single-chain polypeptides containing only one light chain variable domain, single-chain polypeptides containing the three CDRs of the light-chain variable domain, single-chain polypeptides containing only one heavy chain variable region, and single-chain polypeptides containing the three CDRs of the heavy chain variable region.

[0069] “Antigen” refers to proteins that have the ability to generate an immune response in a host. An antigen may be recognized and bound by an antibody. An antigen may originate from within the body or from the external environment.

[0070] “Coding sequence” or “encoding nucleic acid” as used herein may mean refers to the nucleic acid (RNA or DNA molecule) that comprise a nucleotide sequence which encodes an antibody as set forth herein. The coding sequence may further include initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of an individual or mammal to whom the nucleic acid is administered. The coding sequence may further include sequences that encode signal peptides.

[0071] “Complement” or “complementary” as used herein may mean a nucleic acid may mean Watson-Crick (e.g.. A-T / U and C-G) or Hoogsteen base pairing between nucleotides or nucleotide analogs of nucleic acid molecules.

[0072] “Fragment” may mean a polypeptide fragment of an antibody that is function, i.e.. can bind to desired target and have the same intended effect as a full length antibody. A fragment of an antibody may be 100% identical to the full length except missing at least one amino acid from the N and / or C terminal, in each case with or without signal peptides and / or a methionine at position 1. Fragments may comprise 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more. 80% or more, 85% or more, 90% or more, 91% or more, 92% or more, 93% or more. 94% or more, 95% or more, 96% or more. 97% or more, 98% or more, 99% or more percent of the length of the particular full length antibody, excluding any heterologous signal peptide added. The fragment may comprise a fragment of a polypeptide that is 95% or more, 96% or more, 97% or more, 98% or more or 99% or more identical to the antibody and additionally comprise an N terminal methionine or heterologous signal peptide which is not included when calculating percent identity. Fragments may further comprise an N terminal methionine and / or a signal peptide such as an immunoglobulin signal peptide, for example an IgE or IgG signal peptide. The N terminal methionine and / or signal peptide may be linked to a fragment of an antibody. A fragment of a nucleic acid sequence that encodes an antibody may be 100% identical to the full length except missing at least one nucleotide from the 5' and / or 3' end. in each case with or without sequences encoding signal peptides and / or a methionine at position 1. Fragments may comprise 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more. 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more percent of the length of the particular full length coding sequence, excluding any heterologous signal peptide added. The fragment may comprise a fragment that encode a polypeptide that is 95% or more, 96% or more, 97% or more, 98% or more or 99% or more identical to the antibody and additionally optionally comprise sequence encoding an N terminal methionine or heterologous signal peptide which is not included when calculating percent identity. Fragments may further comprise coding sequences for an N terminal methionine and / or a signal peptide such as an immunoglobulin signal peptide, for example an IgE or IgG signal peptide. The coding sequence encoding the N terminal methionine and / or signal peptide may be linked to a fragment of coding sequence.

[0073] “Genetic construct” as used herein refers to the DNA or RNA molecules that comprise a nucleotide sequence which encodes a protein, such as an antibody. The coding sequence includes initiation and termination signals operably linked to regulatory’ elements including a promoter and polyadenylation signal capable of directing expression in the cells of the individual to whom the nucleic acid molecule is administered. As used herein, the term "expressible form" refers to gene constructs that contain the necessary regulatory elements operable linked to a coding sequence that encodes a protein such that when present in the cell of the individual, the coding sequence will be expressed.

[0074] “Identical” or “identity” as used herein in the context of two or more nucleic acids or polypeptide sequences, may mean that the sequences have a specified percentage of residues that are the same over a specified region. The percentage may be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA. thymine (T) and uracil (U) may be considered equivalent. Identity may be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

[0075] “Nucleic acid” or “oligonucleotide” or “polynucleotide” as used herein maymean at least two nucleotides covalently linked together. The depiction of a single strand also defines the sequence of the complementary strand. Thus, a nucleic acid also encompasses the complementary' strand of a depicted single strand. Many variants of a nucleic acid may be used for the same purpose as a given nucleic acid. Thus, a nucleic acid also encompasses substantially identical nucleic acids and complements thereof. A single strand provides a probe that may hybridize to a target sequence under stringent hybridization conditions. Thus, a nucleic acid also encompasses a probe that hybridizes under stringent hybridization conditions.

[0076] Nucleic acids may be single stranded or double stranded, or may contain portions of both double stranded and single stranded sequence. The nucleic acid may be DNA, both genomic and cDNA, RNA, or a hybrid, where the nucleic acid may contain combinations of deoxy ribo- and ribo-nucleotides, and combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine. Nucleic acids may be obtained by chemical synthesis methods or by recombinant methods.

[0077] “Operably linked” as used herein may mean that expression of a gene is under the control of a promoter with which it is spatially connected. A promoter may be positioned 5' (upstream) or 3' (downstream) of a gene under its control. The distance between the promoter and a gene may be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. As is known in the art, variation in this distance may be accommodated without loss of promoter function.

[0078] A “peptide,” “protein,” or “polypeptide” as used herein can mean a linked sequence of amino acids and can be natural, synthetic, or a modification or combination of natural and synthetic.

[0079] “Promoter” as used herein may mean a synthetic or naturally-derived molecule which is capable of conferring, activating or enhancing expression of a nucleic acid in a cell. A promoter may comprise one or more specific transcriptional regulatory sequences to further enhance expression and / or to alter the spatial expression and / or temporal expression of same. A promoter may also comprise distal enhancer or repressor elements, which can be located as much as several thousand base pairs from the start site of transcription. A promoter may be derived from sources including viral, bacterial, fungal, plants, insects, and animals. A promoter may regulate the expression of a gene component constitutively, or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions, or inducing agents. Representative examples of promoters include the bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator-promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV IE promoter, SV40 early promoter or SV 40 late promoter and the CMV IE promoter.

[0080] ■‘Signal peptide” and “leader sequence” are used interchangeably herein and refer to an amino acid sequence that can be linked at the amino terminus of a protein set forth herein. Signal peptides / leader sequences typically direct localization of a protein. Signal peptides / leader sequences used herein preferably facilitate secretion of the protein from the cell in which it is produced. Signal peptides / leader sequences are often cleaved from the remainder of the protein, often referred to as the mature protein, upon secretion from the cell. Signal peptides / leader sequences are linked at the N terminus of the protein.

[0081] “Stringent hybridization conditions” as used herein may mean conditions under which a first nucleic acid sequence (e.g., probe) will hybridize to a second nucleic acid sequence (e.g., target), such as in a complex mixture of nucleic acids. Stringent conditions are sequence dependent and will be different in different circumstances. Stringent conditions may be selected to be about 5-10°C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength pH. The Tmmay be the temperature (under defined ionic strength, pH, and nucleic concentration) at which 50% of the probes complementary to the target hybridize to the target sequence at equilibrium (as the target sequences are present in excess, at Tm, 50% of the probes are occupied at equilibrium). Stringent conditions may be those in which the salt concentration is less than about 1.0 M sodium ion, such as about 0.01- 1.0 M sodium ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes (e.g., about 10-50 nucleotides) and at least about 60°C for long probes (e.g., greater than about 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. For selective or specific hybridization, a positive signal may be at least 2 to 10 times background hybridization. Exemplary stringent hybridization conditions include the following: 50% formamide, 5x SSC. and 1% SDS. incubating at 42°C, or, 5x SSC, 1% SDS. incubating at 65°C. with wash in 0.2x SSC, and 0.1% SDS at 65°C.

[0082] “Subject” and “patient” as used herein interchangeably refers to any vertebrate, including, but not limited to, a mammal (e g., cow, pig, camel, llama, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog, rat, and mouse, a non-human primate (for example, a monkey, such as a cynomolgous or rhesus monkey, chimpanzee, etc) and a human). In some embodiments, the subject may be a human or a non-human. The subject or patient may be undergoing other forms of treatment.

[0083] “Substantially complementary” as used herein may mean that a first sequence is at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the complement of a second sequence over a region of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80. 85, 90, 95, 100 or more nucleotides or amino acids, or that the two sequences hybridize under stringent hybridization conditions.

[0084] “Substantially identical” as used herein may mean that a first and second sequence are at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% over a region of 1, 2, 3, 4, 5, 6. 7, 8, 9. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23. 24. 25. 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100 or more nucleotides or amino acids, or with respect to nucleic acids, if the first sequence is substantially complementary to the complement of the second sequence.

[0085] “Synthetic antibody” as used herein refers to an antibody that is encoded by the recombinant nucleic acid sequence described herein and is generated in a subject.

[0086] “Treatment” or “treating,” as used herein can mean protecting of a subject from a disease through means of preventing, suppressing, repressing, or completely eliminating the disease. Preventing the disease involves administering a vaccine of the present invention to a subject prior to onset of the disease. Suppressing the disease involves administering a vaccine of the present invention to a subject after induction of the disease but before its clinical appearance. Repressing the disease involves administering a vaccine of the present invention to a subject after clinical appearance of the disease.

[0087] “Variant” used herein with respect to a nucleic acid may mean (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a nucleic acid that is substantially identical to a referenced nucleic acid or the complement thereof; or (iv) a nucleic acid that hybridizes under stringent conditions to the referenced nucleic acid, complement thereof, or a sequences substantially identical thereto.

[0088] “Variant’' with respect to a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity. Variant may also mean a protein with an amino acid sequence that is substantially identical to a referenced protein with an amino acid sequence that retains at least one biological activity. A conservative substitution of an amino acid, i.e., replacing an amino acid with a different amino acid of similar properties (e.g., hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes can be identified, in part, by considering the hydropathic index of amino acids, as understood in the art. Kyte et al., J. Mol. Biol. 157: 105-132 (1982). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydropathic indexes can be substituted and still retain protein function. In one aspect, amino acids having hydropathic indexes of ±2 are substituted. The hydrophilicity of amino acids can also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide, a useful measure that has been reported to correlate well with antigenicity and immunogenicity. U.S. Patent No. 4,554,101, incorporated fully herein by reference. Substitution of amino acids having similar hydrophilicity values can result in peptides retaining biological activity, for example immunogenicity, as is understood in the art. Substitutions may be performed with amino acids having hydrophilicity values within ±2 of each other. Both the hyrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties.

[0089] A variant may be a nucleic acid sequence that is substantially identical over the full length of the full gene sequence or a fragment thereof. The nucleic acid sequence may be 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the full length of the gene sequence or a fragment thereof. A variant may be an amino acid sequence that is substantially identical over the full length of the amino acid sequence or fragment thereof. The amino acid sequence may be 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the full length of the amino acid sequence or a fragment thereof.

[0090] ■‘Vector” as used herein may mean a nucleic acid sequence containing an origin of replication. A vector may be a plasmid, bacteriophage, bacterial artificial chromosome or yeast artificial chromosome. A vector may be a DNA or RNA vector. A vector may be either a self-replicating extrachromosomal vector or a vector which integrates into a host genome.

[0091] For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1. 6.2, 6.3, 6.4, 6.5. 6.6, 6.7, 6.8, 6.9. and 7.0 are explicitly contemplated.

[0092] Composition

[0093] In some embodiments, the invention provides compositions comprising molecules that bind to an influenza antigen, including, but not limited to, influenza a virus hemagglutinin (HA) or anti influenza a virus neuraminidase (NA). In some embodiments, the molecule that binds HA or NA is an antibody.

[0094] The invention relates to a composition comprising a recombinant nucleic acid sequence encoding an antibody, a fragment thereof, a variant thereof, or a combination thereof. The composition, when administered to a subject in need thereof, can result in the generation of a synthetic antibody in the subject. The synthetic antibody can bind a target molecule (i.e., an antigen) present in the subject. Such binding can neutralize the antigen, block recognition of the antigen by another molecule, for example, a protein or nucleic acid, and elicit or induce an immune response to the antigen.

[0095] In one embodiment, the composition comprises a nucleotide sequence encoding a synthetic antibody. In one embodiment, the composition comprises a nucleic acid molecule comprising a first nucleotide sequence encoding a first synthetic antibody and a second nucleotide sequence encoding a second synthetic antibody. In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding a cleavage domain.

[0096] In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding an antibody to HA. In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding an antibody to NA.

[0097] In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding a variable heavy chain region and a nucleotide sequence encoding a variable light chain region of an anti -HA antibody.

[0098] In one embodiment, the invention provides a composition comprising a first nucleic acid molecule comprising a nucleotide sequence encoding a variable heavy chain region of an anti-HA antibody and a second nucleic acid molecule comprising a nucleotide sequence encoding a variable light chain region of an anti-HA antibody.

[0099] In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding a variable heavy chain region and a nucleotide sequence encoding a variable light chain region of an anti-NA antibody.

[0100] In one embodiment, the invention provides a composition comprising a first nucleic acid molecule comprising a nucleotide sequence encoding a variable heavy chain region of an anti-NA antibody and a second nucleic acid molecule comprising a nucleotide sequence encoding a variable light chain region of an anti-NA antibody.

[0101] Antibodies, including HA or NA binding protein fragments, of the present invention include, in certain embodiments, antibody amino acid sequences disclosed herein encoded by any suitable polynucleotide, or any isolated or formulated antibody. Further, antibodies of the present disclosure comprise antibodies having the structural and / or functional features of anti-HA or anti-NA antibodies described herein. In one embodiment, the anti-HA antibody binds HA and, thereby partially or substantially alters at least one biological activity of the HA protein (e.g., receptor binding activity). In one embodiment, the anti-NA antibody binds NA and. thereby partially or substantially alters at least one biological activity of the NA protein (e.g.. receptor binding activity).

[0102] In some embodiments, the invention provides a composition comprising at least one influenza A antigen or a nucleic acid molecule encoding an antigen. In some embodiments, the influenza A antigen is an immunogen. In some embodiments, the immunogen is an HA immunogen or a nucleoprotein (NP) immunogen. In some embodiments, the immunogen is a consensus antigen. Antibodies

[0103] In some embodiments, the invention includes compositions comprising an antibody that specifically binds to HA (e.g., binding portion of an antibody). In one embodiment, the anti -HA antibody is a polyclonal antibody. In another embodiment, the anti- HA antibody is a monoclonal antibody. In some embodiments, the anti-HA antibody is a chimeric antibody. In further embodiments, the anti-HA antibody is a humanized antibody.

[0104] In some embodiments, the invention includes compositions comprising an antibody that specifically binds to NA (e.g., binding portion of an antibody). In one embodiment, the anti-MA antibody is a polyclonal antibody. In another embodiment, the anti-NA antibody is a monoclonal antibody. In some embodiments, the anti-NA antibody is a chimeric antibody. In further embodiments, the anti-NA antibody is a humanized antibody.

[0105] The binding portion of an antibody comprises one or more fragments of an antibody that retain the ability to specifically bind to binding partner molecule (e.g., HA or NA). It has been shown that the binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term '‘binding portion” of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL. VH, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CHI domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single chain antibodies are also intended to be encompassed within the term '‘binding portion” of an antibody. These antibody fragments are obtained using conventional techniques known to those with skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies. Binding portions can be produced by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact immunoglobulins. The antibody may comprise a heavy chain and a light chain complementarity determining region (“CDR”) set, respectively interposed between a heavy chain and a light chain framework (“FR”) set which provide support to the CDRs and define the spatial relationship of the CDRs relative to each other. The CDR set may contain three hypervariable regions of a heavy or light chain V region. Proceeding from the N-terminus of a heavy or light chain, these regions are denoted as “CDR1.” “CDR2,” and “CDR3,” respectively. An antigen-binding site, therefore, may include six CDRs, comprising the CDR set from each of a heavy and a light chain V region.

[0106] The antibody can be an immunoglobulin (Ig). The Ig can be, for example, IgA, IgM, IgD. IgE, and IgG. The immunoglobulin can include the heavy chain polypeptide and the light chain polypeptide. The heavy chain polypeptide of the immunoglobulin can include a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region. The light chain polypeptide of the immunoglobulin can include a VL region and CL region.

[0107] The antibody can be a polyclonal or monoclonal antibody. The antibody can be a chimeric antibody, a single chain antibody, an affinity matured antibody, a human antibody, a humanized antibody, or a fully human antibody. The humanized antibody can be an antibody from a non-human species that binds the desired antigen having one or more complementarity determining regions (CDRs) from the non-human species and framework regions from a human immunoglobulin molecule.

[0108] As described herein, the antibody can be generated in the subject upon administration of the composition to the subject. The antibody may have a half-life within the subject. In some embodiments, the antibody may be modified to extend or shorten its half-life within the subject. Such modifications are described below in more detail.

[0109] The antibody can be defucosylated.

[0110] The antibody may be modified to reduce or prevent antibody-dependent enhancement (ADE) of disease associated with the antigen as described in more detail below.

[0111] In one embodiment, the HA antibody comprises a heavy chain comprising an amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24. In one embodiment, the HA antibody comprises a light chain comprising an amino acid sequence as set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SE ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46. Given that certain of the monoclonal antibodies can bind to H A. the VH and VL sequences can be “mixed and matched” to create other anti-HA binding molecules of this disclosure. Binding of such “mixed and matched” antibodies can be tested using standard binding assays known in the art (e.g., immunoblot etc.). In some embodiments, when VH and VL chains are mixed and matched, a VH sequence from a particular VH / VL pairing is replaced with a structurally similar VH sequence. Likewise, preferably a VL sequence from a particular VH / VL pairing is replaced with a structurally similar VL sequence.

[0112] Accordingly, in one embodiment, this disclosure provides an isolated monoclonal antibody , or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO:4; and (b) a light chain amino acid sequence of SEQ ID NO:26.

[0113] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO:6; and (b) a light chain amino acid sequence of SEQ ID NO:42.

[0114] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO: 8; and (b) a light chain amino acid sequence of SEQ ID NO:28.

[0115] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO: 10: and (b) a light chain amino acid sequence of SEQ ID NO:30.

[0116] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy7chain amino acid sequence of SEQ ID NO: 12; and (b) a light chain amino acid sequence of SEQ ID NO:32.

[0117] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO: 14; and (b) a light chain amino acid sequence of SEQ ID NO:34.

[0118] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO: 16: and (b) a light chain amino acid sequence of SEQ ID NO:36.

[0119] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO: 18; and (b) a light chain amino acid sequence of SEQ ID NO:38. In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO:20; and (b) a light chain amino acid sequence of SEQ ID NO:40.

[0120] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO:22: and (b) a light chain amino acid sequence of SEQ ID NO:44.

[0121] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO:24; and (b) a light chain amino acid sequence of SEQ ID NO:46.

[0122] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising an amino acid sequence of SEQ ID NO:2.

[0123] In one embodiment, anti -HA antibody comprises a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity7to an amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10. SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16. SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24. In one embodiment, anti-HA antibody comprises a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30. SEQ ID NO: 32. SEQ ID NO: 34. SEQ ID NO: 36, SE ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46.

[0124] In one embodiment, the NA antibody comprises a heavy chain comprising an amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51. In one embodiment, the NA antibody comprises a light chain comprising an amino acid sequence as set forth in SEQ ID NO: 49. SEQ ID NO: 53.

[0125] Accordingly, in one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy chain amino acid sequence of SEQ ID NO:47; and (b) a light chain amino acid sequence of SEQ ID NO:49.

[0126] In one embodiment, this disclosure provides an isolated monoclonal antibody, or binding portion thereof comprising: (a) a heavy7chain amino acid sequence of SEQ ID NO:51; and (b) a light chain amino acid sequence of SEQ ID NO:53.

[0127] In one embodiment, anti -NA antibody comprises a heavy7chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51. In one embodiment, anti -HA antibody comprises a light chain amino acid sequence having at least 60%, 70%, 80%. 90%. 95%. 96%. 97%. 98%. or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 49, SEQ ID NO: 53.

[0128] Extension of Antibody Half-Life

[0129] The antibody may be modified to extend or shorten the half-life of the antibody in the subject. The modification may extend or shorten the half-life of the antibody in the serum of the subject.

[0130] The modification may be present in a constant region of the antibody. The modification may be one or more ammo acid substitutions in a constant region of the antibody that extend the half-life of the antibody as compared to a half-life of an antibody not containing the one or more amino acid substitutions. The modification may be one or more amino acid substitutions in the CH2 domain of the antibody that extend the half-life of the antibody as compared to a half-life of an antibody not containing the one or more amino acid substitutions.

[0131] In some embodiments, the one or more amino acid substitutions in the constant region may include replacing a methionine residue in the constant region with a tyrosine residue, a serine residue in the constant region with a threonine residue, a threonine residue in the constant region with a glutamate residue, or any combination thereof, thereby extending the half-life of the antibody.

[0132] In other embodiments, the one or more amino acid substitutions in the constant region may include replacing a methionine residue in the CH2 domain with a tyrosine residue, a serine residue in the CH2 domain with a threonine residue, a threonine residue in the CH2 domain with a glutamate residue, or any combination thereof, thereby extending the half-life of the antibody.

[0133] Defucosylation

[0134] The antibody may be an antibody that is not fucosylated (i.e., a defucosylated antibody or a non-fucosylated antibody), a fragment thereof, a variant thereof, or a combination thereof. Fucosylation includes the addition of the sugar fucose to a molecule, for example, the attachment of fucose to N-glycans. O-glycans and glycolipids. Accordingly, in a defucosylated antibody, fucose is not attached to the carbohydrate chains of the constant region. In turn, this lack of fucosylation may improve FcyRIIIa binding and antibody- directed cellular cytotoxic (ADCC) activity by the antibody as compared to the fucosylated antibody. Therefore, in some embodiments, the non-fucosylated antibody may exhibit increased ADCC activity as compared to the fucosylated antibody.

[0135] The antibody may be modified so as to prevent or inhibit fucosylation of the antibody. In some embodiments, such a modified antibody may exhibit increased ADCC activity as compared to the unmodified antibody. The modification may be in the heavy chain, light chain, or a combination thereof. The modification may be one or more amino acid substitutions in the heavy chain, one or more amino acid substitutions in the light chain, or a combination thereof.

[0136] Reduced ADE Response

[0137] The antibody may be modified to reduce or prevent antibody-dependent enhancement (ADE) of disease associated with the antigen, but still neutralize the antigen.

[0138] In some embodiments, the antibody may be modified to include one or more amino acid substitutions that reduce or prevent binding of the antibody to FcyRla. The one or more amino acid substitutions may be in the constant region of the antibody.

[0139] Bispecific Antibody

[0140] The antibody may be a bispecific antibody, a fragment thereof, a variant thereof, or a combination thereof. The bispecific antibody can bind or react with two antigens, for example, two of the antigens described below in more detail. The bispecific antibody can be comprised of fragments of two of the antibodies described herein, thereby allowing the bispecific antibody to bind or react with two desired target molecules, which may include the antigen, which is described below in more detail, a ligand, including a ligand for a receptor, a receptor, including a ligand-binding site on the receptor, a ligand-receptor complex, and a marker.

[0141] The invention provides novel bispecific antibodies comprising a first antigenbinding site that specifically binds to a first target and a second antigen-binding site that specifically binds to a second target, with particularly advantageous properties such as producibility, stability, binding affinity-, biological activity, specific targeting of certain T cells, targeting efficiency and reduced toxicity. In some instances, there are bispecific antibodies, wherein the bispecific antibody binds to the first target with high affinity and to the second target with low affinity. In other instances, there are bispecific antibodies, wherein the bispecific antibody binds to the first target with low affinity and to the second target with high affinity. In other instances, there are bispecific antibodies, wherein the bispecific antibody binds to the first target with a desired affinity and to the second target with a desired affinity.

[0142] In one embodiment, the bispecific antibody is a bivalent antibody comprising a) a first light chain and a first heavy chain of an antibody specifically binding to a first antigen, and b) a second light chain and a second heavy chain of an antibody specifically binding to a second antigen.

[0143] A bispecific antibody molecule according to the invention may have two binding sites of any desired specificity. In some embodiments, the binding site included in the Fab fragment is a binding site specific for an HA or NA antigen. In some embodiments, the binding site included in the single chain Fv fragment is a binding site specific for an HA or NA antigen.

[0144] Bifunctional Antibody

[0145] The antibody may be a bifunctional antibody, a fragment thereof, a variant thereof, or a combination thereof. The bifunctional antibody can bind or react with the antigen described below. The bifunctional antibody can also be modified to impart an additional functionality to the antibody beyond recognition of and binding to the antigen. Such a modification can include, but is not limited to, coupling to factor H or a fragment thereof. Factor H is a soluble regulator of complement activation and thus, may contribute to an immune response via complement-mediated lysis (CML).

[0146] Antigen

[0147] The synthetic antibody may be directed to the antigen or fragment or variant thereof. The antigen can be a nucleic acid sequence, an amino acid sequence, a polysaccharide or a combination thereof. The nucleic acid sequence can be DNA, RNA, cDNA, a variant thereof, a fragment thereof, or a combination thereof. The amino acid sequence can be a protein, a peptide, a variant thereof, a fragment thereof, or a combination thereof. The poly saccharide can be a nucleic acid encoded poly saccharide.

[0148] The antigen can be from a virus. The antigen can be associated with viral infection. In one embodiment, the antigen can be associated with influenza infection. In one embodiment, the antigen can be an HA or NA antigen. In one embodiment, the antigen can be a fragment of an HA or NA antigen.

[0149] In one embodiment, a synthetic antibody of the invention targets two or more antigens. In one embodiment, at least one antigen of a bispecific antibody is selected from the antigens described herein. In one embodiment, the two or more antigens are selected from the antigens described herein.

[0150] Viral Antigens

[0151] The viral antigen can be a viral antigen or fragment or variant thereof. The virus can be a disease-causing virus. The virus can be an influenza virus.

[0152] The antigen may be an influenza viral antigen, or fragment thereof, or variant thereof. The influenza antigen can be from a factor that allows the virus to replicate, infect or survive. Factors that allow an influenza virus to infect or survive include, but are not limited to, hemagglutinin.

[0153] Generation of Synthetic Antibodies In Vitro and Ex Vivo

[0154] In one embodiment, one or more synthetic antibody is generated in vitro or ex vivo. For example, in one embodiment, a nucleic acid encoding a synthetic antibody can be introduced and expressed in an in vitro or ex vivo cell. Methods of introducing and expressing genes into a cell are known in the art. In the context of an expression vector, the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art. For example, the expression vector can be transferred into a host cell by physical, chemical, or biological means.

[0155] Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and / or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al. (2012, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). A preferred method for the introduction of a polynucleotide into a host cell is calcium phosphate transfection.

[0156] Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. Other viral vectors can be derived from lentivirus, poxviruses, herpes simplex virus I, adenoviruses and adeno-associated viruses, and the like. See. for example, U.S. Pat. Nos. 5,350,674 and 5,585,362.

[0157] Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a deli \ ery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle).

[0158] In the case where a non-viral delivery' system is utilized, an exemplary' delivery vehicle is a liposome or lipid nanoparticle. The use of lipid formulations is contemplated for the introduction of the nucleic acids into a host cell (in vitro, ex vivo or in vivo). In another aspect, the nucleic acid may be associated with a lipid. The nucleic acid associated wi th a lipid may be encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the oligonucleotide, entrapped in a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid. Lipid, lipid / DNA or lipid / expression vector associated compositions are not limited to any particular structure in solution. For example, they may be present in a bilayer structure, as micelles, or with a “collapsed’’ structure. They may also simply be interspersed in a solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty' substances which may be naturally occurring or synthetic lipids. For example, lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty' acids, alcohols, amines, amino alcohols, and aldehydes.

[0159] Recombinant Nucleic Acid Sequence- Antibodies

[0160] In some aspects, the composition can comprise a recombinant nucleic acid sequence encoding an antibody, a fragment thereof, a variant thereof, or a combination thereof. The recombinant nucleic acid sequence can be a heterologous nucleic acid sequence. The recombinant nucleic acid sequence can include at least one heterologous nucleic acid sequence or one or more heterologous nucleic acid sequences.

[0161] The recombinant nucleic acid sequence can be an optimized nucleic acid sequence. Such optimization can increase or alter the immunogenicity of the antibody. Optimization can also improve transcription and / or translation. Optimization can include one or more of the following: low GC content leader sequence to increase transcription; mRNA stability and codon optimization; addition of a kozak sequence (e.g., GCC ACC) for increased translation; addition of an immunoglobulin (Ig) leader sequence encoding a signal peptide; and eliminating to the extent possible cis-acting sequence motifs (i.e., internal TATA boxes).

[0162] In one embodiment, the anti-HA antibody comprises a DMAb encoded by a single plasmid which comprises sequences encoding both the heavy and light chains of the antibody. In one embodiment, the DMAb comprises a nucleic acid molecule encoding the amino acid sequence of SEQ ID NO:2, an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:2, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:2.

[0163] In one embodiment, the invention relates to a nucleotide sequence encoding an anti-HA antibody comprises a nucleic acid sequence encoding a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10. SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16. SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises a codon optimized nucleic acid sequence encoding a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of an amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10. SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16. SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24, wherein the sequence encodes each of the CDRs of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NOTO, SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO:16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO 22, or SEQ ID NO 24.

[0164] In one embodiment, the invention relates to a nucleotide sequence encoding a heavy chain of an anti-HA antibody comprising a nucleotide sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%. 98%. or 99% identity to SEQ ID NO:3, SEQ ID NO:5, SEQ ID N0:7. SEQ ID NO:9. SEQ ID NO: 11, SEQ ID NO: 13. SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises a codon optimized nucleic acid sequence comprising at least 60%, 70%. 80%. 90%. 95%. 96%. 97%, 98%, or 99% of the full length of a nucleotide sequence as set forth in SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23, wherein the sequence encodes each of the CDRs of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14. DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22. or SEQ ID NO: 24.

[0165] In one embodiment, the invention relates to a nucleotide sequence encoding an anti-HA antibody comprises a nucleic acid sequence encoding a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30. SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises a codon optimized nucleic acid sequence encoding a fragment comprising at least 60%, 70%, 80%, 90%, 95%. 96%, 97%. 98%. or 99% of the full length of an amino acid sequence as set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SE ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46, wherein the sequence encodes each of the CDRs of SEQ ID NO: 26. SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SE ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44. or SEQ ID NO: 46.

[0166] In one embodiment, the nucleic acid sequence encoding a light chain amino acid sequence has at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25, SEQ ID NO:27. SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33. SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43, SEQ ID NO: 45. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises a codon optimized nucleic acid sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of a nucleotide sequence as set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37. SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43, SEQ ID NO: 45, wherein the sequence encodes each of the CDRs of SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30. SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SE ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46.

[0167] In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14. DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22. or SEQ ID NO: 24.

[0168] In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a nucleotide sequence as set forth in SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO:15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain comprising a nucleotide sequence as set forth in SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO:17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17. SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23.

[0169] In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 26. SEQ ID NO: 28. SEQ ID NO: 30. SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain amino acid sequence as set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a fragment comprising at least 60%. 70%. 80%. 90%. 95%. 96%. 97%. 98%. or 99% of the full length of SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46.

[0170] In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a nucleotide sequence as set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO: 41. SEQ ID NO:43, SEQ ID NO: 45, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain comprising a nucleotide sequence as set forth in SEQ ID NO:25, SEQ ID NO:27. SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35. SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43, SEQ ID NO: 45. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain comprising a nucleotide sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35. SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43. SEQ ID NO: 45.

[0171] In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain, a light chain or a combination thereof of an antibody of the invention. In one embodiment, the nucleic acid molecule encodes SEQ ID NO: 2. an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:2, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:2. In one embodiment, the nucleotide sequence encoding an anti-HA antibody comprises an RNA sequence transcribed from a DNA sequence comprising SEQ ID NO: 1, a nucleotide sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a DNA sequence comprising SEQ ID NO: 1, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO: 1.

[0172] In one embodiment, the invention relates to a combination of a first nucleic acid molecule encoding a heavy chain of an anti-HA antibody, and a second nucleic acid molecule encoding a light chain of an anti-HA antibody. In one embodiment, the first nucleic acid molecule is a first plasmid comprising a nucleotide sequence encoding a heavy chain of an anti-HA antibody and the second nucleic acid molecule is a second plasmid encoding a light chain of an anti-HA antibody.

[0173] In one embodiment, the first nucleic acid molecule encoding a heavy chain of an anti-HA antibody encodes SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14. DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22. or SEQ ID NO:24. or an amino acid sequence having at least 60%. 70%. 80%. 90%. 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO:24, or a fragment comprising at least 60%. 70%. 80%. 90%. 95%. 96%. 97%. 98%. or 99% of the full length of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, DEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, or SEQ ID NO: 24.

[0174] In one embodiment, the first nucleic acid molecule encoding a heavy chain of an anti-HA antibody composes SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13. SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21. or SEQ ID NO:23. or a nucleotide sequence having at least 60%. 70%. 80%. 90%. 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID N0:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%. 96%. 97%. 98%. or 99% of the full length of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 1 1, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23.

[0175] In one embodiment, the second nucleic acid molecule encoding a light chain of an anti-HA antibody encodes SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, or SEQ ID NO: 46, an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36. SEQ ID NO: 38. SEQ ID NO: 40. SEQ ID NO: 42. SEQ ID NO: 44, or SEQ ID NO: 46 or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44. or SEQ ID NO: 46.

[0176] In one embodiment, the second nucleic acid molecule encoding a light chain of an anti-HA antibody comprises SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO 37, SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43, SEQ ID NO: 45, or a nucleotide sequence having at least 60%, 70%, 80%, 90%. 95%. 96%. 97%. 98%. or 99% identity to SEQ ID NO:25, SEQ ID NO:27. SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO 35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43, SEQ ID NO: 45, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:25, SEQ ID NO:27. SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37. SEQ ID NO:39, SEQ ID NO: 41, SEQ ID NO:43, SEQ ID NO: 45.

[0177] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:3. encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:25, encoding a light chain amino acid sequence.

[0178] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:5, encoding a heavy7chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:41. encoding a light chain amino acid sequence.

[0179] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:7, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:27, encoding a light chain amino acid sequence.

[0180] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:9. encoding a heavy7chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:29, encoding a light chain amino acid sequence.

[0181] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 11, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 31, encoding a light chain amino acid sequence.

[0182] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 13, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:33. encoding a light chain amino acid sequence.

[0183] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 15, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:35. encoding a light chain amino acid sequence.

[0184] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 17, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:37, encoding a light chain amino acid sequence. In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 19, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:39, encoding a light chain amino acid sequence.

[0185] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:21, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:43, encoding a light chain amino acid sequence.

[0186] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:23, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:45, encoding a light chain amino acid sequence.

[0187] In one embodiment, the nucleic acid molecule is a single plasmid encoding a heavy and light chain of a DMAb. In one embodiment, the nucleic acid molecule encodes SEQ ID NO:2. an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identify to an amino acid sequence as set forth in SEQ ID NO:2, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:2. In one embodiment, the nucleic acid molecule is a single plasmid comprising a nucleotide sequence of SEQ ID NO: 1. a nucleotide sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identify to SEQ ID NO: 1, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO: 1.

[0188] In one embodiment, the anti-NA antibody comprises a DMAb encoded by a single plasmid which comprises sequences encoding both the heavy and light chains of the antibody. In one embodiment, the DMAb comprises a nucleic acid molecule encoding the amino acid sequences of SEQ ID NO:47 and SEQ ID NO:49, or SEQ ID NO:51 and SEQ ID NO: 53, an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%. or 99% identify to an amino acid sequence as set forth in SEQ ID NO:47 and SEQ ID NO:49, or SEQ ID NO:51 and SEQ ID NO: 53 or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:47 and SEQ ID NO:49, or SEQ ID NO:51 and SEQ ID NO: 53.

[0189] In one embodiment, the invention relates to a nucleotide sequence encoding an anti-NA antibody comprises a nucleic acid sequence encoding a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51. In one embodiment, the nucleotide sequence encoding an anti-HNA antibody comprises a codon optimized nucleic acid sequence encoding a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of an amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51, wherein the sequence encodes each of the CDRs of SEQ ID NO:47 or SEQ ID NO:51.

[0190] In one embodiment, the invention relates to a nucleotide sequence encoding a heavy chain of an anti-NA antibody comprising a nucleotide sequence having at least 60%, 70%, 80%, 90%, 95%, 96%. 97%. 98%. or 99% identity to SEQ ID NO:48 or SEQ ID NO: 52. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises a codon optimized nucleic acid sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of a nucleotide sequence as set forth in SEQ ID NO:48 or SEQ ID NO:52, wherein the sequence encodes each of the CDRs of SEQ ID NO:47 or SEQ ID NO:51.

[0191] In one embodiment, the invention relates to a nucleotide sequence encoding an anti-NA antibody comprises a nucleic acid sequence encoding a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 49 or SEQ ID NO: 53. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises a codon optimized nucleic acid sequence encoding a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of an amino acid sequence as set forth in SEQ ID NO: 49 or SEQ ID NO: 53, wherein the sequence encodes each of the CDRs of SEQ ID NO: 49 or SEQ ID NO: 53.

[0192] In one embodiment, the nucleic acid sequence encoding a light chain amino acid sequence has at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 50 or SEQ ID NO: 54. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises a codon optimized nucleic acid sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of a nucleotide sequence as set forth in SEQ ID NO: 50 or SEQ ID NO: 54, wherein the sequence encodes each of the CDRs of SEQ ID NO: 49 or SEQ ID NO: 53.

[0193] In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:47 or SEQ ID NO:51.

[0194] In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a nucleotide sequence as set forth in SEQ ID NO:48 or SEQ ID NO:52, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a heavy’ chain comprising a nucleotide sequence as set forth in SEQ ID NO:48 or SEQ ID NO:52. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%. 97%. 98%. or 99% of the full length of SEQ ID NO:48 or SEQ ID NO:52.

[0195] In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity7to an amino acid sequence as set forth in SEQ ID NO: 49 or SEQ ID NO: 53. or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain amino acid sequence as set forth in SEQ ID NO: 49 or SEQ ID NO: 53. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a fragment comprising at least 60%. 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO: 49 or SEQ ID NO: 53.

[0196] In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a nucleotide sequence as set forth in SEQ ID NO: 50 or SEQ ID NO: 54, or a combination thereof. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain comprising a nucleotide sequence as set forth in SEQ ID NO: 50 or SEQ ID NO: 54. In one embodiment, the nucleotide sequence encoding an anti-NA antibody comprises an RNA sequence transcribed from a DNA sequence encoding a light chain comprising a nucleotide sequence comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO: 50 or SEQ ID NO: 54.

[0197] In one embodiment, the invention relates to a combination of a first nucleic acid molecule encoding a heavy chain of an anti-NA antibody, and a second nucleic acid molecule encoding a light chain of an anti-NA antibody. In one embodiment, the first nucleic acid molecule is a first plasmid comprising a nucleotide sequence encoding a heavy chain of an anti-NA antibody and the second nucleic acid molecule is a second plasmid encoding a light chain of an anti-NA antibody.

[0198] In one embodiment, the first nucleic acid molecule encoding a heavy chain of an anti-NA antibody encodes SEQ ID NO:47 or SEQ ID NO:51, or an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO:47 or SEQ ID NO:51, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:47 or SEQ ID NO:51.

[0199] In one embodiment, the first nucleic acid molecule encoding a heavy chain of an anti-NA antibody comprises SEQ ID NO:48 or SEQ ID NO:52, or a nucleotide sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:48 or SEQ ID NO:52, or a fragment comprising at least 60%. 70%. 80%. 90%. 95%. 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:48 or SEQ ID NO:52.

[0200] In one embodiment, the second nucleic acid molecule encoding a light chain of an anti-NA antibody encodes SEQ ID NO:49 or SEQ ID NO:53, an amino acid sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence as set forth in SEQ ID NO: 49 or SEQ ID NO: 53 or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:49 or SEQ ID NO:53.

[0201] In one embodiment, the second nucleic acid molecule encoding a light chain of an anti-NA antibody comprises SEQ ID NO:50 or SEQ ID NO:54, or a nucleotide sequence having at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:50 or SEQ ID NO:54, or a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO:50 or SEQ ID NO:54.

[0202] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:48, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:50, encoding a light chain amino acid sequence.

[0203] In one embodiment, the invention relates to a combination of a first nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO:52, encoding a heavy chain amino acid sequence, and a second nucleic acid molecule comprising a nucleotide sequence of SEQ ID NO: 54, encoding a light chain amino acid sequence.

[0204] Heavv Chain Polypeptide

[0205] The recombinant nucleic acid sequence construct can include the heterologous nucleic acid encoding the heavy chain polypeptide, a fragment thereof, a variant thereof, or a combination thereof. The heavy chain polypeptide can include a variable heavy chain (VH) region and / or at least one constant heavy chain (CH) region. The at least one constant heavy chain region can include a constant heavy chain region 1 (CHI), a constant heavy chain region 2 (CH2), and a constant heavy chain region 3 (CH3), and / or a hinge region.

[0206] In some embodiments, the heavy chain polypeptide can include a VH region and a CHI region. In other embodiments, the heavy chain polypeptide can include a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region.

[0207] The heavy’ chain polypeptide can include a complementarity determining region C‘CDR”) set. The CDR set can contain three hypervariable regions of the VH region. Proceeding from N-terminus of the heavy chain polypeptide, these CDRs are denoted “CDR1,” CDR2.' and “CDR3,’’ respectively. CDR1, CDR2, and CDR3 of the heavy chain polypeptide can contribute to binding or recognition of the antigen.

[0208] Light Chain Polypeptide

[0209] The recombinant nucleic acid sequence construct can include the heterologous nucleic acid sequence encoding the light chain polypeptide, a fragment thereof, a variant thereof, or a combination thereof. The light chain polypeptide can include a variable light chain (VL) region and / or a constant light chain (CL) region. The light chain polypeptide can include a complementarity determining region C'CDR”) set. The CDR set can contain three hypervariable regions of the VL region. Proceeding from N-terminus of the light chain polypeptide, these CDRs are denoted '‘CDR1,” “CDR2,” and “CDR3,” respectively. CDR1, CDR2, and CDR3 of the light chain polypeptide can contribute to binding or recognition of the antigen.

[0210] Promoter

[0211] The recombinant nucleic acid sequence construct can include one or more promoters. The one or more promoters may be any promoter that is capable of driving gene expression and regulating gene expression. Such a promoter is a cis-acting sequence element required for transcription via a DNA dependent RNA polymerase. Selection of the promoter used to direct gene expression depends on the particular application. The promoter may be positioned about the same distance from the transcription start in the recombinant nucleic acid sequence construct as it is from the transcription start site in its natural setting. However, variation in this distance may be accommodated without loss of promoter function.

[0212] The promoter may be operably linked to the heterologous nucleic acid sequence encoding the heavy chain polypeptide and / or light chain polypeptide. The promoter may be a promoter shown effective for expression in eukaryotic cells. The promoter operably linked to the coding sequence may be a CMV promoter, a promoter from simian virus 40 (SV40), such as SV40 early promoter and SV40 later promoter, a mouse mammary tumor virus (MMTV) promoter, a human immunodeficiency virus (HIV) promoter such as the bovine immunodeficiency virus (BIV) long terminal repeat (LTR) promoter, a Moloney virus promoter, an avian leukosis virus (ALV) promoter, a cytomegalovirus (CMV) promoter such as the CMV immediate early promoter, Epstein Ban virus (EBV) promoter, or a Rous sarcoma virus (RSV) promoter. The promoter may also be a promoter from a human gene such as human actin, human myosin, human hemoglobin, human muscle creatine, human polyhedrin, or human metalothionein.

[0213] The promoter can be a constitutive promoter or an inducible promoter, which initiates transcription only when the host cell is exposed to some particular external stimulus. In the case of a multicellular organism, the promoter can also be specific to a particular tissue or organ or stage of development. The promoter may also be a tissue specific promoter, such as a muscle or skin specific promoter, natural or synthetic. Examples of such promoters are described in US patent application publication no. US20040175727, the contents of which are incorporated herein in its entirety.

[0214] The promoter can be associated with an enhancer. The enhancer can be located upstream of the coding sequence. The enhancer may be human actin, human myosin, human hemoglobin, human muscle creatine or a viral enhancer such as one from CMV, FMDV, RSV or EBV. Polynucleotide function enhances are described in U.S. Patent Nos. 5,593,972, 5,962,428, and W094 / 016737, the contents of each are fully incorporated by reference.

[0215] Intron

[0216] The recombinant nucleic acid sequence construct can include one or more introns. Each intron can include functional splice donor and acceptor sites. The intron can include an enhancer of splicing. The intron can include one or more signals required for efficient splicing.

[0217] Transcription Termination Region

[0218] The recombinant nucleic acid sequence construct can include one or more transcription termination regions. The transcription termination region can be downstream of the coding sequence to provide for efficient termination. The transcription termination region can be obtained from the same gene as the promoter described above or can be obtained from one or more different genes.

[0219] Initiation Codon

[0220] The recombinant nucleic acid sequence construct can include one or more initiation codons. The initiation codon can be located upstream of the coding sequence. The initiation codon can be in frame with the coding sequence. The initiation codon can be associated with one or more signals required for efficient translation initiation, for example, but not limited to, a ribosome binding site.

[0221] Termination Codon

[0222] The recombinant nucleic acid sequence construct can include one or more termination or stop codons. The termination codon can be downstream of the coding sequence. The termination codon can be in frame with the coding sequence. The termination codon can be associated with one or more signals required for efficient translation termination.

[0223] Polyadenylation Signal

[0224] The recombinant nucleic acid sequence construct can include one or more polyadenylation signals. The polyadenylation signal can include one or more signals required for efficient poly adenylation of the transcript. The poly adenylation signal can be positioned downstream of the coding sequence. The poly adenylation signal may be a SV40 polyadenylation signal, LTR polyadenylation signal, bovine grow th hormone (bGH) polyadenylation signal, human growth hormone (hGH) polyadenylation signal, or human fl- globin polyadenylation signal. The SV40 polyadenylation signal may be a polyadenylation signal from a pCEP4 plasmid (Invitrogen, San Diego, CA).

[0225] Leader Sequence

[0226] The recombinant nucleic acid sequence construct can include one or more leader sequences. The leader sequence can encode a signal peptide. The signal peptide can be an immunoglobulin (Ig) signal peptide, for example, but not limited to, an IgG signal peptide and a IgE signal peptide.

[0227] Expression of antibodies from the Recombinant Nucleic Acid Sequence Construct

[0228] The recombinant nucleic acid sequence construct can include, amongst the one or more components, the heterologous nucleic acid sequence encoding the heavy chain polypeptide and / or the heterologous nucleic acid sequence encoding the light chain polypeptide. Accordingly, the recombinant nucleic acid sequence construct can facilitate expression of the heavy chain polypeptide and / or the light chain polypeptide.

[0229] Upon expression, for example, but not limited to, in a cell, organism, or mammal, the heavy chain polypeptide and the light chain polypeptide can assemble into the synthetic antibody. In particular, the heavy chain polypeptide and the light chain polypeptide can interact with one another such that assembly results in the synthetic antibody being capable of binding the antigen. In other embodiments, the heavy chain polypeptide and the light chain polypeptide can interact with one another such that assembly results in the synthetic antibody being more immunogenic as compared to an antibody not assembled as described herein. In still other embodiments, the heavy chain polypeptide and the light chain polypeptide can interact with one another such that assembly results in the synthetic antibodybeing capable of eliciting or inducing an immune response against the antigen.

[0230] Vector

[0231] The recombinant nucleic acid sequence construct described above can be placed in one or more vectors. The one or more vectors can contain an origin of replication. The one or more vectors can be a plasmid, bacteriophage, bacterial artificial chromosome or yeast artificial chromosome. The one or more vectors can be either a self-replication extra chromosomal vector, or a vector which integrates into a host genome.

[0232] The one or more vectors can be a heterologous expression construct, which is generally a plasmid that is used to introduce a specific gene into a target cell. Once the expression vector is inside the cell, the antibody- or antibody fragment that is encoded by' the recombinant nucleic acid sequence construct is produced by the cellular-transcription and translation machinery- ribosomal complexes. The one or more vectors can express large amounts of stable messenger RNA, and therefore proteins.

[0233] Expression Vector

[0234] The one or more vectors can be a circular plasmid or a linear nucleic acid.

[0235] The circular plasmid and linear nucleic acid are capable of directing expression of a particular nucleotide sequence in an appropriate subject cell. The one or more vectors comprising the recombinant nucleic acid sequence construct may' be chimeric, meaning that at least one of its components is heterologous with respect to at least one of its other components.

[0236] Plasmid

[0237] The one or more vectors can be a plasmid. The plasmid may be useful for transfecting cells with the recombinant nucleic acid sequence construct. The plasmid may be useful for introducing the recombinant nucleic acid sequence construct into the subject. The plasmid may also comprise a regulatory sequence, which may- be well suited for gene expression in a cell into which the plasmid is administered.

[0238] The plasmid may- also comprise a mammalian origin of replication in order to maintain the plasmid extrachromosomally and produce multiple copies of the plasmid in a cell. The plasmid may be pVAXl. pCEP4 or pREP4 from Invitrogen (San Diego, CA), which may comprise the Epstein Barr virus origin of replication and nuclear antigen EBNA-1 coding region, which may produce high copy episomal replication without integration. The backbone of the plasmid may be pAV0242. The plasmid may be a replication defective adenovirus type 5 (Ad5) plasmid.

[0239] The plasmid may be pSE420 (Invitrogen, San Diego, Calif), which may be used for protein production in Escherichia coli (E.coli). The plasmid may also be p YES2 (Invitrogen, San Diego, Calif.), which may be used for protein production in Saccharomyces cerevisiae strains of yeast. The plasmid may also be of the MAXBAC™ complete baculovirus expression system (Invitrogen, San Diego, Calif), which may be used for protein production in insect cells. The plasmid may also be pcDNAI or pcDNA3 (Invitrogen, San Diego, Calif), which may be used for protein production in mammalian cells such as Chinese hamster ovary (CHO) cells.

[0240] Circular and Linear Vector

[0241] The one or more vectors may be circular plasmid, which may transform a target cell by integration into the cellular genome or exist extrachromosomally (e.g., autonomous replicating plasmid with an origin of replication). The vector can be pVAX, pcDNA3.0, or provax, or any other expression vector capable of expressing the antibody or antibody fragment encoded by the recombinant nucleic acid sequence construct.

[0242] Also provided herein is a linear nucleic acid, or linear expression cassette (‘ LEC’’), that is capable of being efficiently delivered to a subject via electroporation and expressing the antibody or antibody fragment encoded by the recombinant nucleic acid sequence construct. The LEC may be any linear DNA devoid of any phosphate backbone. The LEC may not contain any antibiotic resistance genes and / or a phosphate backbone. The LEC may not contain other nucleic acid sequences unrelated to the desired gene expression.

[0243] The LEC may be derived from any plasmid capable of being linearized. The plasmid may be capable of expressing the antibody or antibody fragment encoded by the recombinant nucleic acid sequence construct. The plasmid can be pNP (Puerto Rico / 34) or pM2 (New Caledonia / 99). The plasmid may be WLV009, pVAX, pcDNA3.0, or provax, or any other expression vector capable of expressing the antibody or antibody fragment encoded by the recombinant nucleic acid sequence construct.

[0244] The LEC can be pcrM2. The LEC can be pcrNP. pcrNP and pcrMR can be derived from pNP (Puerto Rico / 34) and pM2 (New Caledonia / 99), respectively. Viral Vectors

[0245] In one embodiment, viral vectors are provided herein which are capable of delivering a nucleic acid of the invention to a cell. The expression vector may be provided to a cell in the form of a viral vector. Viral vector technology7is well know n in the art and is described, for example, in Sambrook et al. (2001), and in Ausubel et al. (1997), and in other virology and molecular biology manuals. Viruses, which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers. (See. e g., WO 01 / 96584; WO 01 / 29058; and U.S. Pat. No. 6,326,193. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. Other viral vectors can be derived from lentivirus, poxviruses, herpes simplex virus I, adenoviruses and adeno- associated viruses, and the like. See, for example, U.S. Pat. Nos. 5,350.674 and 5,585,362.

[0246] Nanoparticle Formulations

[0247] In one embodiment, the composition of the invention may comprise a nanoparticle, including but not limited to a lipid nanoparticle (LNP), comprising an HA or NA antibody of the invention, or a LNP comprising a nucleic acid encoding an HA or NA antibody of the invention. In some embodiments, the composition comprises or encodes all or part of an HA or NA antigen binding molecule of the invention, or an immunogenically functional equivalent thereof. In some embodiments, the composition comprises an mRNA molecule that encodes all or part of an HA or NA antigen binding molecule of the invention.

[0248] In one embodiment, the immunogenic composition of the invention may comprise a composition comprising a combination of HA or NA antibodies of the invention, or a LNP comprising one or more nucleic acid molecules encoding a combination of HA or NA antibodies of the invention. In one embodiment, the immunogenic composition of the invention may comprise a composition comprising a combination of LNP, wherein the combination of LNP comprises one or more nucleic acid molecules encoding a combination of HA or NA antibodies of the invention.

[0249] In one embodiment, the LNP comprises or encapsulates an RNA molecule encoding at least one amino acid sequence of SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18. SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28. SEQ ID NQ:30, SEQ ID NO:32. SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38. SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51,or SEQ ID NO:53 or a fragment or variant thereof.

[0250] In one embodiment, the LNP comprises or encapsulates an RNA molecule comprising a nucleotide sequence corresponding to. or transcribed from SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO 31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39. SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, or SEQ ID NO:54 or a fragment or variant thereof.

[0251] In one embodiment, the invention provides a combination of LNPs comprising or encapsulating a combination of RNA molecules encoding an anti -HA antibody. In some embodiments the combination of LNPs comprises a first nucleic acid molecule encoding a heavy chain of an anti-HA antibody encodes SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, or SEQ ID NO:23, and a second nucleic acid molecule encoding a light chain of an anti-HA antibody encodes SEQ ID NO:25, SEQ ID NO:27. SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO 33, SEQ ID NO:35, SEQ ID NO:37. SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, or SEQ ID NO:45, or a fragment thereof comprising at least the variable region thereof.

[0252] In one embodiment, the invention provides a combination of LNPs comprising or encapsulating a combination of RNA molecules encoding an anti -NA antibody. In some embodiments the combination of LNPs comprises a first nucleic acid molecule encoding a heavy chain of an anti-NA antibody encodes SEQ ID NO:47 or SEQ ID NO:51, and a second nucleic acid molecule encoding a light chain of an anti-NA antibody encodes SEQ ID NO:50 or SEQ ID NO: 53, or a fragment thereof comprising at least the variable region thereof.

[0253] In one embodiment, the invention relates to a combination of LNPs comprising or encapsulating a combination of at least two RNA molecules encoding the combination of the heavy chain and light chain of the synthetic antibody of the invention or fragments or variants thereof. In one embodiment, the composition further comprises one or more additional immunostimulatory agents. Immunostimulatory agents include, but are not limited to, an additional antigen or antigen binding molecule, an immunomodulator, or an adjuvant.

[0254] Arrangement of the Recombinant Nucleic Acid Sequence Construct

[0255] As described above, the recombinant nucleic acid sequence can include one or more recombinant nucleic acid sequence constructs, in which each recombinant nucleic acid sequence construct can include one or more components. The one or more components are described in detail above. The one or more components, when included in the recombinant nucleic acid sequence construct, can be arranged in any order relative to one another. In some embodiments, the one or more components can be arranged in the recombinant nucleic acid sequence construct as described below.

[0256] Arrangement 1

[0257] In one arrangement, a first recombinant nucleic acid sequence construct can include the heterologous nucleic acid sequence encoding the heavy chain polypeptide and a second recombinant nucleic acid sequence construct can include the heterologous nucleic acid sequence encoding the light chain polypeptide. The first recombinant nucleic acid sequence construct can be placed in a vector. The second recombinant nucleic acid sequence construct can be placed in a second or separate vector. Placement of the recombinant nucleic acid sequence construct into the vector is described in more detail below.

[0258] The first recombinant nucleic acid sequence construct can also include the promoter, intron, transcription termination region, initiation codon, termination codon, and / or polyadenylation signal. The first recombinant nucleic acid sequence construct can further include the leader sequence, in which the leader sequence is located upstream (or 5’) of the heterologous nucleic acid sequence encoding the heavy chain polypeptide. Accordingly, the signal peptide encoded by the leader sequence can be linked by a peptide bond to the heavy chain polypeptide.

[0259] The second recombinant nucleic acid sequence construct can also include the promoter, initiation codon, termination codon, and polyadenylation signal. The second recombinant nucleic acid sequence construct can further include the leader sequence, in which the leader sequence is located upstream (or 5’) of the heterologous nucleic acid sequence encoding the light chain polypeptide. Accordingly, the signal peptide encoded by the leader sequence can be linked by a peptide bond to the light chain polypeptide.

[0260] Accordingly, one example of arrangement 1 can include the first vector (and thus first recombinant nucleic acid sequence construct) encoding the heavy’ chain polypeptide that includes VH and CHI, and the second vector (and thus second recombinant nucleic acid sequence construct) encoding the light chain polypeptide that includes VL and CL. A second example of arrangement 1 can include the first vector (and thus first recombinant nucleic acid sequence construct) encoding the heavy chain polypeptide that includes VH, CHI, hinge region, CH2, and CH3, and the second vector (and thus second recombinant nucleic acid sequence construct) encoding the light chain polypeptide that includes VL and CL.

[0261] Arrangement 2

[0262] In a second arrangement, the recombinant nucleic acid sequence construct can include the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide. The heterologous nucleic acid sequence encoding the heavy chain polypeptide can be positioned upstream (or 5’) of the heterologous nucleic acid sequence encoding the light chain polypeptide. Alternatively, the heterologous nucleic acid sequence encoding the light chain polypeptide can be positioned upstream (or 5?) of the heterologous nucleic acid sequence encoding the heavy chain polypeptide.

[0263] The recombinant nucleic acid sequence construct can be placed in the vector as described in more detail below.

[0264] The recombinant nucleic acid sequence construct can include the heterologous nucleic acid sequence encoding the protease cleavage site and / or the linker sequence. If included in the recombinant nucleic acid sequence construct, the heterologous nucleic acid sequence encoding the protease cleavage site can be positioned between the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide. Accordingly, the protease cleavage site allows for separation of the heavy chain polypeptide and the light chain polypeptide into distinct polypeptides upon expression. In other embodiments, if the linker sequence is included in the recombinant nucleic acid sequence construct, then the linker sequence can be positioned between the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide. The recombinant nucleic acid sequence construct can also include the promoter, intron, transcription termination region, initiation codon, termination codon, and / or polyadenylation signal. The recombinant nucleic acid sequence construct can include one or more promoters. The recombinant nucleic acid sequence construct can include two promoters such that one promoter can be associated with the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the second promoter can be associated with the heterologous nucleic acid sequence encoding the light chain polypeptide. In still other embodiments, the recombinant nucleic acid sequence construct can include one promoter that is associated with the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide.

[0265] The recombinant nucleic acid sequence construct can further include two leader sequences, in which a first leader sequence is located upstream (or 5’) of the heterologous nucleic acid sequence encoding the heavy chain polypeptide and a second leader sequence is located upstream (or 5‘) of the heterologous nucleic acid sequence encoding the light chain polypeptide. Accordingly, a first signal peptide encoded by the first leader sequence can be linked by a peptide bond to the heavy chain polypeptide and a second signal peptide encoded by the second leader sequence can be linked by a peptide bond to the light chain polypeptide.

[0266] Accordingly, one example of arrangement 2 can include the vector (and thus recombinant nucleic acid sequence construct) encoding the heavy chain polypeptide that includes VH and CHI, and the light chain polypeptide that includes VL and CL, in which the linker sequence is positioned between the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide.

[0267] A second example of arrangement of 2 can include the vector (and thus recombinant nucleic acid sequence construct) encoding the heavy chain polypeptide that includes VH and CHI, and the light chain polypeptide that includes VL and CL, in which the heterologous nucleic acid sequence encoding the protease cleavage site is positioned between the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide.

[0268] A third example of arrangement 2 can include the vector (and thus recombinant nucleic acid sequence construct) encoding the heavy chain polypeptide that includes VH, CHI, hinge region, CH2, and CH3, and the light chain polypeptide that includes VL and CL. in which the linker sequence is positioned between the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide.

[0269] A forth example of arrangement of 2 can include the vector (and thus recombinant nucleic acid sequence construct) encoding the heavy chain polypeptide that includes VH. CHI. hinge region, CH2, and CH3, and the light chain polypeptide that includes VL and CL, in which the heterologous nucleic acid sequence encoding the protease cleavage site is positioned between the heterologous nucleic acid sequence encoding the heavy chain polypeptide and the heterologous nucleic acid sequence encoding the light chain polypeptide.

[0270] Antigens and Immunogens

[0271] In some embodiments, the invention provides a composition comprising at least one influenza A antigen or a nucleic acid molecule encoding an antigen. In some embodiments, the influenza A antigen is an immunogen. In some embodiments, the immunogen is an HA immunogen or a nucleoprotein (NP) immunogen. In some embodiments, the immunogen is a consensus antigen. In some embodiments, the immunogen is a consensus antigen derived from H1N1 or H3N2.

[0272] In some embodiments, the invention relates to immunogens that recapitulate the antigenicity of influenza A nucleoprotein (NP) or hemagglutinin (HA) protein. In some embodiments, the immunogen is suitable for vaccination strategies to stimulate an immune response, in a subject. In one embodiment, the immunogen is able to elicit detectable antibody responses and heterologous neutralizing antibodies against influenza A infection.

[0273] In one embodiment, the composition comprises a peptide comprising an amino acid sequence capable of expressing one or more NP or HA antigens in the subject and a pharmaceutically acceptable excipient.

[0274] In one embodiment, the composition comprises nucleotide sequences capable of expressing an NP or HA antigen in the subject and a pharmaceutically acceptable excipient. In one embodiment, the nucleic acid molecule comprises a promoter operably linked to a coding sequence that encodes an NP or HA antigen. In some embodiments, the composition comprises nucleotide sequences capable of expressing a self-assembling ferritin nanoparticle decorated with an NP or HA antigen. In one embodiment, the immunogen comprises a polypeptide comprising amino acid sequence having at least 60%. 70%. 80%. 90%. 95%. 96%. 97%. 98%. or 99% identity to an amino acid sequence as set forth in SEQ ID NO:56, SEQ ID NO:58, or SEQ ID NO: 60. In some embodiments, the immunogen comprises a fragment comprising at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the full length of SEQ ID NO: 56, SEQ ID NO:58, or SEQ ID NO:60.

[0275] In another aspect, this disclosure provides immunogen polypeptides that are multimerized. However, the invention is not limited to any means of multimerizing. Rather, the invention includes any means of multimerizing including but is not limited to multimerizing on a nanoparticle, on a virus-like particle (VLP) and the likes. Virus-like particles, or retrovirus-like particles, in the context of the present disclosure, are membrane- surrounded structures comprising viral envelope proteins embedded within the membrane of the host cell in which they are produced, and preferably, additional viral core proteins in the VLPs. These VLPs do not contain intact viral nucleic acid, and they are non- infectious. Desirably, there is sufficient envelope protein on the surface of the VLP so that when a VLP preparation is formulated into an immunogenic composition and administered to an animal or human, an immune response (cell-mediated or humoral) is raised.

[0276] In another aspect, this disclosure provides a protein complex comprising at least one above-described immunogen polypeptide multimerized via covalent or non-covalent bonding / interaction (e.g., van der Waals interactions). For example, two or more immunogen polypeptides may be cross-linked by one or more cross-linkers. Crosslinkers are reagents having reactive ends to specific functional groups (e.g. , primary7amines or sulfhydryls) on proteins or other molecules. Crosslinkers are capable of joining two or more molecules by a covalent bond. Crosslinkers include but are not limited to amine- to-amine crosslinkers (e.g.. disuccinimidyl suberate(DSS)), amine-to-sulfhydryl crosslinkers (e.g. , N-g- maleimidobutyryl- oxysuccinimide ester (GMBS)), carboxyl-to-amine crosslinkers (e.g., dicyclohexylcarbodiimide (DCC)), sulfhydiyl-to-carbohydrate crosslinkers (e.g., N-b- maleimidopropionic acid hydrazide (BMPH)), sulfhydryl-to-sulfhydryl crosslinkers (e.g., 1,4- bismaieimidobutane (BMB)), photoreactive crosslinkers (e.g. , N-5-azido-2- nitrobenzoyloxy succinimide (ANB-NOS)), chemo selective ligation crosslinkers (e.g., NHS- PEG4-Azide).

[0277] In one embodiment, the present invention provides an immunogenic composition comprising one or more nucleic acid molecules that are capable of generating in a mammal an immune response against an influenza A antigen. The present invention also provides isolated nucleic acid molecules that are capable of generating in a mammal an immune response against an influenza antigen. In one embodiment, the immunogenic composition comprises amino acid sequences for an influenza antigen having the amino acid sequence set forth in v SEQ ID NO:56, SEQ ID NO:58, or SEQ ID NO:60 SEQ ID NO:56, SEQ ID NO:58, or SEQ ID NO:60 or fragments or variants thereof.

[0278] In one embodiment, the invention provides compositions comprising a nucleic acid molecule comprising a nucleotide sequence that encodes an influenza A antigen. In one embodiment, the nucleic acid molecule comprises a nucleic acid sequence SEQ ID NO: 56, SEQ ID NO:58, or SEQ ID NO:60, or fragments or variants thereof. In one embodiment, a nucleotide sequence which encodes an HIV antigen is provided as SEQ ID NO:55, SEQ ID NO:57, or SEQ ID NO:59, or fragments or variants thereof.

[0279] Compositions that comprise one or more nucleotide sequence that encode an influenza antigen may be on a single plasmid. In one embodiment, a composition comprises a single plasmid that encodes an influenza antigen under a single promoter.

[0280] In one embodiment, an influenza antigen is operably linked to one or more regulatory elements. In one embodiment, a regulatory element is a leader sequence. In one embodiment, a regulatory' element is a start codon. In one embodiment, a regulatory element is at least one stop codon.

[0281] When taken up by a cell, the DNA plasmids can remain in the cell as separate genetic material. Alternatively, RNA may' be administered to the cell. It is also contemplated to provide a genetic construct as a linear minichromosome including a centromere, telomeres and an origin of replication. Genetic constructs include regulatory’ elements necessary for gene expression of a nucleic acid molecule. The elements include: a promoter, an initiation codon, a stop codon, and a polyadenylation signal. In addition, enhancers are often required for gene expression of the sequence that encodes the target protein or the immunomodulating protein. It is necessary that these elements be operable linked to the sequence that encodes the desired proteins and that the regulatory elements are operably in the individual to whom they are administered. Such genetic constructs may be therefore be recombinant nucleic acid molecules.

[0282] The recombinant nucleic acid molecule can include one or more recombinant nucleotide sequence constructs. The recombinant nucleotide sequence construct can include a heterologous nucleotide sequence that encodes a viral antigen, a fragment thereof, a variant thereof, or a combination thereof.

[0283] The recombinant nucleotide sequence construct can include one or more leader sequences. The leader sequence can encode a signal peptide. The signal peptide can be an immunoglobulin (Ig) signal peptide, for example, but not limited to, an IgG signal peptide and a IgE signal peptide. In some embodiments, nucleic acid constructs may be provided in which the coding sequences for the proteins described herein are linked to IgE leader peptide, or such IgE leader is removed. In some embodiments, proteins described herein are linked to IgE signal peptide, or such IgE leader is removed.

[0284] The one or more vectors can be a plasmid. The plasmid may be useful for transfecting cells with the recombinant nucleotide sequence construct. The plasmid may be useful for introducing the recombinant nucleotide sequence construct into the subject. The plasmid may also comprise a regulatory7sequence, which may be well suited for gene expression in a cell into which the plasmid is administered.

[0285] The plasmid may also comprise a mammalian origin of replication in order to maintain the plasmid extrachromosomally and produce multiple copies of the plasmid in a cell. The plasmid may be pVAXl, pCEP4 or pREP4 from Invitrogen (San Diego, CA), which may comprise the Epstein Barr virus origin of replication and nuclear antigen EBNA-1 coding region, which may produce high copy episomal replication without integration. The backbone of the plasmid may be pAV0242. The plasmid may be a replication defective adenovirus ty pe 5 (Ad5) plasmid.

[0286] The plasmid may be pSE420 (Invitrogen, San Diego, Calif), which may be used for protein production in Escherichia coli (E.coli). The plasmid may also be pYES2 (Invitrogen, San Diego, Calif.), which may be used for protein production in Saccharomyces cerevisiae strains of yeast. The plasmid may7also be of the MAXBAC™ complete baculovirus expression system (Invitrogen, San Diego, Calif), which may be used for protein production in insect cells. The plasmid may also be pcDNAI or pcDNA3 (Invitrogen, San Diego, Calif), which may be used for protein production in mammalian cells such as Chinese hamster ovary (CHO) cells.

[0287] In one embodiment, the composition of the invention may comprise a nanoparticle, including but not limited to a lipid nanoparticle (LNP), comprising an NP or HA antigen of the invention, or a LNP comprising a nucleic acid encoding an NP or HA antigen of the invention. In some embodiments, the composition comprises or encodes all or part of an NP or HA antigen of the invention, or an immunogenically functional equivalent thereof. In some embodiments, the composition comprises an mRNA molecule that encodes all or part of an NP or HA antigen of the invention.

[0288] In one embodiment, the immunogenic composition of the invention may comprise a composition comprising a combination of NP or HA antigens of the invention, or a LNP comprising one or more nucleic acid molecules encoding a combination of NP or HA antigens of the invention. In one embodiment, the immunogenic composition of the invention may comprise a composition comprising a combination of LNP, wherein the combination of LNP comprises one or more nucleic acid molecules encoding a combination of NP or HA antigens of the invention.

[0289] In one embodiment, the LNP comprises or encapsulates an RNA molecule encoding at least one amino acid sequence of SEQ ID NO:56, SEQ ID NO: 58, or SEQ ID NO: 60, or a fragment or variant thereof. In one embodiment, the LNP comprises or encapsulates an RNA molecule comprising a nucleotide sequence corresponding to, or transcribed from SEQ ID NO:55, SEQ ID NO:57. or SEQ ID NO:59. or a fragment or variant thereof.

[0290] Excipients and Other Components of the Composition

[0291] The composition may further comprise a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient can be functional molecules such as vehicles, carriers, or diluents. The pharmaceutically acceptable excipient can be a transfection facilitating agent, which can include surface active agents, such as immune- stimulating complexes (ISCOMS), Freunds incomplete adjuvant, LPS analog including monophosphoryl lipid A, muramyl peptides, quinone analogs, vesicles such as squalene and squalene, hyaluronic acid, lipids, liposomes, calcium ions, viral proteins, polyanions, poly cations, or nanoparticles, or other known transfection facilitating agents.

[0292] The transfection facilitating agent is a poly anion, poly cation, including poly- L-glutamate (LGS), or lipid. The transfection facilitating agent is poly-L-glutamate, and the poly-L-glutamate may be present in the composition at a concentration less than 6 mg / ml. The transfection facilitating agent may also include surface active agents such as immune- stimulating complexes (ISCOMS), Freunds incomplete adjuvant, LPS analog including monophosphoryl lipid A, muramyl peptides, quinone analogs and vesicles such as squalene and squalene, and hyaluronic acid may also be used administered in conjunction with the composition. The composition may also include a transfection facilitating agent such as lipids, liposomes, including lecithin liposomes or other liposomes known in the art, as a DNA-liposome mixture (see for example W09324640). calcium ions, viral proteins, polyanions, poly cations, or nanoparticles, or other known transfection facilitating agents. The transfection facilitating agent is a poly anion, poly cation, including poly-L-glutamate (LGS), or lipid. Concentration of the transfection agent in the vaccine is less than 4 mg / ml, less than 2 mg / ml, less than 1 mg / ml, less than 0.750 mg / ml, less than 0.500 mg / ml, less than 0.250 mg / ml, less than 0.100 mg / ml, less than 0.050 mg / ml, or less than 0.010 mg / ml.

[0293] The composition may further comprise a genetic facilitator agent as described in U.S. Serial No. 021,579 filed April 1, 1994, which is fully incorporated by reference.

[0294] In some embodiments of the present invention, the composition further includes an adjuvant. In some embodiments, the adjuvant is selected from the group consisting of: IL- 12 alpha-interferon, gamma-interferon, platelet derived growth factor (PDGF), TNFa, TNFp, GM-CSF, epidermal grow th factor (EGF), cutaneous T cell-attracting chemokine (CTACK), epithelial thymus-expressed chemokine (TECK), mucosae-associated epithelial chemokine (MEC), IL- 15. MHC, CD80,CD86 including IL- 15 having the signal sequence deleted and optionally including the signal peptide from IgE. Other genes which may be useful adjuvants include those encoding: MCP-1, MIP-l-alpha, MIP-lp, IL-8, RANTES, L-selectin, P-selectin. E-selectin, CD34, GlyCAM-1, MadCAM-1, LFA-1, VLA- 1, Mac-1, pl50.95, PECAM, ICAM-1. ICAM-2. ICAM-3, CD2, LFA-3, M-CSF, G-CSF, IL- 4, mutant forms of IL- 18, CD40, CD40L, vascular growth factor, fibroblast growth factor, IL-7, nerve growth factor, vascular endothelial growth factor, Fas, TNF receptor, Fit, Apo-1, p55, WSL-1, DR3, TRAMP, Apo-3, AIR, LARD, NGRF, DR4, DR5, KILLER, TRAIL-R2, TRICK2. DR6, Caspase ICE, Fos, c-jun, Sp-1, Ap-1, Ap-2, p38, p65Rel, MyD88, IRAK, TRAF6, IkB. Inactive NIK, SAP K, SAP-1. JNK. interferon response genes, NFkB, Bax, TRAIL, TRAILrec, TRAILrecDRC5, TRAIL-R3, TRAIL-R4, RANK, RANK LIGAND, 0x40, 0x40 LIGAND, NKG2D, MICA, MICB, NKG2A, NKG2B, NKG2C, NKG2E, NKG2F, TAPI, TAP2 and functional fragments thereof. In some preferred embodiments, the adjuvant is IL-12.

[0295] The composition may comprise DNA at quantities of from about 1 nanogram to 100 milligrams; about 1 microgram to about 10 milligrams; or preferably about 0. 1 microgram to about 10 milligrams; or more preferably about 1 milligram to about 2 milligram. In some preferred embodiments, composition according to the present invention comprises about 5 nanogram to about 1000 micrograms of DNA. In some preferred embodiments, composition can contain about 10 nanograms to about 800 micrograms of DNA. In some preferred embodiments, the composition can contain about 0. 1 to about 500 micrograms of DNA. In some preferred embodiments, the composition can contain about 1 to about 350 micrograms of DNA. In some preferred embodiments, the composition can contain about 25 to about 250 micrograms, from about 100 to about 200 microgram, from about 1 nanogram to 100 milligrams; from about 1 microgram to about 10 milligrams; from about 0.1 microgram to about 10 milligrams; from about 1 milligram to about 2 milligram, from about 5 nanogram to about 1000 micrograms, from about 10 nanograms to about 800 micrograms, from about 0.1 to about 500 micrograms, from about 1 to about 350 micrograms, from about 25 to about 250 micrograms, from about 100 to about 200 microgram of DNA.

[0296] The composition can be formulated according to the mode of administration to be used. An injectable pharmaceutical composition can be sterile, pyrogen free and particulate free. An isotonic formulation or solution can be used. Additives for isotonicity can include sodium chloride, dextrose, mannitol, sorbitol, and lactose. The composition can comprise a vasoconstriction agent. The isotonic solutions can include phosphate buffered saline. The composition can further comprise stabilizers including gelatin and albumin. The stabilizers can allow the formulation to be stable at room or ambient temperature for extended periods of time, including LGS or polycations or poly anions.

[0297] Methods of Delivery of the Composition

[0298] The present invention also relates to methods of delivering the composition to the subject in need thereof. The method of delivery can include, administering the composition to the subject. The mammal receiving delivery of the composition may be human, primate, non-human primate, cow, cattle, sheep, goat, antelope, bison, water buffalo, bison, bovids, deer, hedgehogs, elephants, llama, alpaca, mice, rats, and chicken.

[0299] The composition may be administered by different routes including orally, parenterally, sublingually, transdermally, rectally, transmucosally, topically, via inhalation, via buccal administration, intrapleurally, intravenous, intraarterial, intraperitoneal, subcutaneous, intramuscular, intranasal intrathecal, and intraarticular or combinations thereof. For veterinary use, the composition may be administered as a suitably acceptable formulation in accordance with normal veterinary practice. The veterinarian can readily determine the dosing regimen and route of administration that is most appropriate for a particular animal. The composition may be administered by traditional syringes, needleless injection devices, “microprojectile bombardment gone guns”, or other physical methods such as electroporation (“EP”), “hydrodynamic method”, or ultrasound.

[0300] Method of Treatment

[0301] Also provided herein is a method of treating, protecting against, and / or preventing disease in a subject in need thereof by administering a synthetic antibody or nucleic acid molecule encoding the synthetic antibody to the subject, and / or administering an immunogen or nucleic acid encoding the immunogen to the subject. The method can include administering the composition to the subject. Administration of the composition to the subject can be done using the method of delivery described above.

[0302] In certain embodiments, the invention provides a method of treating protecting against, and / or preventing a influenza. In certain embodiments, the invention provides a method of treating protecting against, and / or preventing a bacterial infection. In one embodiment, the method treats, protects against, and / or prevents influenza A

[0303] The synthetic antibody can bind to or react with the antigen. Such binding can neutralize an antigen, block recognition of the antigen by another molecule, for example, a protein or nucleic acid, and elicit or induce an immune response to the antigen, thereby treating, protecting against, and / or preventing the disease associated with the antigen in the subject.

[0304] The composition dose can be between 1 pg to 10 mg active component / kg body weight / time, and can be 20 pg to 10 mg component / kg body weight / time. The composition can be administered every 1, 2. 3, 4, 5, 6, 7, 8, 9. 10, 11, 12, 13, 14, 15, 16, 17, 18. 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30. or 31 days. The number of composition doses for effective treatment can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0305] The composition of the invention can treat, prevent and / or protect against any disease, disorder, or condition associated with a bacterial or viral activity. In certain embodiments, the composition can treat, prevent, and or / protect against bacterial or viral infection. In certain embodiments, the composition can treat, prevent, and or / protect against influenza.

[0306] The synthetic antibody and / or antigen can treat, prevent, and / or protect against disease in the subject administered the composition. The synthetic antibody by binding the antigen can treat, prevent, and / or protect against disease in the subject administered the composition. The synthetic antibody can promote survival of the disease in the subject administered the composition. The antigen by eliciting an immune response can treat, prevent, and / or protect against disease in the subject administered the composition. The anbtigen can promote survival of the disease in the subject administered the composition.

[0307] The synthetic antibody can provide at least about 50%, 55%, 60%, 65%, 70%, 75%. 80%. 85%. 90%. 95%. or 100% survival of the disease in the subject administered the composition. In other embodiments, the synthetic antibody can provide at least about 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, or 80% survival of the disease in the subject administered the composition.

[0308] The antigen can provide at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% survival of the disease in the subject administered the composition. In other embodiments, the antigen can provide at least about 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, or 80% survival of the disease in the subj ect administered the composition.

[0309] The composition can result in the generation of the synthetic antibody in the subject within at least about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 20 hours, 25 hours, 30 hours, 35 hours, 40 hours, 45 hours, 50 hours, or 60 hours of administration of the composition to the subject. The composition can result in generation of the synthetic antibody in the subject within at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, or 10 days of administration of the composition to the subject. The composition can result in generation of the synthetic antibody in the subject within about 1 hour to about 6 days, about 1 hour to about 5 days, about 1 hour to about 4 days, about 1 hour to about 3 days, about 1 hour to about 2 days, about 1 hour to about 1 day, about 1 hour to about 72 hours, about 1 hour to about 60 hours, about 1 hour to about 48 hours, about 1 hour to about 36 hours, about 1 hour to about 24 hours, about 1 hour to about 12 hours, or about 1 hour to about 6 hours of administration of the composition to the subject.

[0310] The composition, when administered to the subject in need thereof, can result in the generation of the synthetic antibody in the subject more quickly than the generation of an endogenous antibody in a subject who is administered an antigen to induce a humoral immune response. The composition can result in the generation of the synthetic antibody at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days. 9 days, or 10 days before the generation of the endogenous antibody in the subject who was administered an antigen to induce a humoral immune response.

[0311] The composition of the present invention can have features required of effective compositions such as being safe so that the composition does not cause illness or death; being protective against illness; and providing ease of administration, few side effects, biological stability and low cost per dose. The method of delivering the antigen (i.e. the DNA / plasmid vaccine or vaccination) may be provided to induce a therapeutic and prophylactic immune response. The vaccination process may generate in the mammal an immune response against the antigen. The vaccine may be delivered to an individual to modulate the activity of the mammal’s immune system and enhance the immune response. The delivery of the vaccine may be the transfection of the consensus antigen as a nucleic acid molecule that is expressed in the cell and delivered to the surface of the cell upon which the immune system recognized and induces a cellular, humoral, or cellular and humoral response. The delivery of the vaccine may be used to induce or elicit and immune response in mammals against the antigen by administering to the mammals the vaccine as discussed above.

[0312] The DNA vaccine and the antibody (i.e. the DMAb) may be administered at the same time or at different times. In one embodiment, the DNA vaccine and the DMAb are administered simultaneously. In one embodiment, the DNA vaccine is administered before the DMAb. In one embodiment, the DMAb is administered before the DNA vaccine.

[0313] In certain embodiments, the DNA vaccine is administered 1 or more days, 2 or more days, 3 or more days, 4 or more days, 5 or more days, 6 or more days, 7 or more days, 8 or more days, 9 or more days, 10 or more days, 11 or more days, 12 or more days, 13 or more days, or 14 or more days after the DMAb is administered. In certain embodiments, the DNA vaccine is administered 1 or more weeks, 2 or more weeks. 3 or more weeks. 4 or more weeks, 5 or more weeks, 6 or more weeks, 7 or more weeks, 8 or more weeks, 9 or more weeks, or 10 or more weeks after the DMAb is administered. In certain embodiments, the DNA vaccine is administered 1 or more months, 2 or more months, 3 or more months, 4 or more months, 5 or more months. 6 or more months. 7 or more months, 8 or more months, 9 or more months, 10 or more months, 11 or more months, or 12 or more months after the DMAb is administered.

[0314] In certain embodiments, the DMAb is administered 1 or more days, 2 or more days, 3 or more days, 4 or more days, 5 or more days. 6 or more days, 7 or more days, 8 or more days, 9 or more days, 10 or more days, 11 or more days, 12 or more days, 13 or more days, or 14 or more days after the DNA vaccine is administered. In certain embodiments, the DMAb is administered 1 or more weeks, 2 or more weeks, 3 or more weeks, 4 or more weeks, 5 or more weeks, 6 or more weeks, 7 or more weeks, 8 or more weeks, 9 or more weeks, or 10 or more weeks after the DNA vaccine is administered. In certain embodiments, the DMAb is administered 1 or more months, 2 or more months, 3 or more months, 4 or more months, 5 or more months, 6 or more months. 7 or more months, 8 or more months, 9 or more months, 10 or more months, 11 or more months, or 12 or more months after the DNA vaccine is administered.

[0315] In certain embodiments, the DMAb and DNA vaccine are administered once. In certain embodiments, the DMAb and / or the DNA vaccine are administered more than once. In certain embodiments, administration of the DMAb and DNA vaccine provides a persistent and systemic immune response.

[0316] Use in Combination with Antibiotics

[0317] The present invention also provides a method of treating, protecting against, and / or preventing disease in a subject in need thereof by administering a combination of the synthetic antibody, or nucleic acid molecule encoding the synthetic antibody, and one or more additional thereapeutic agent. In some embodiments, the additional therapeutic agent is an antibiotic agent.

[0318] The synthetic antibody and an antibiotic agent may be administered using any suitable method such that a combination of the synthetic antibody and antibiotic agent are both present in the subj ect. In one embodiment, the method may comprise administration of a first composition comprising a synthetic antibody of the invention by any of the methods described in detail above and administration of a second composition comprising an antibiotic agent less than 1, less than 2, less than 3, less than 4, less than 5, less than 6, less than 7, less than 8, less than 9 or less than 10 days following administration of the synthetic antibody. In one embodiment, the method may comprise administration of a first composition comprising a synthetic antibody of the invention by any of the methods described in detail above and administration of a second composition comprising an antibiotic agent more than 1, more than 2, more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or more than 10 days following administration of the synthetic antibody. In one embodiment, the method may comprise administration of a first composition comprising an antibiotic agent and administration of a second composition comprising a synthetic antibody of the invention by any of the methods described in detail above less than 1, less than 2. less than 3. less than 4. less than 5. less than 6. less than 7, less than 8, less than 9 or less than 10 days following administration of the antibiotic agent. In one embodiment, the method may comprise administration of a first composition comprising an antibiotic agent and administration of a second composition comprising a synthetic antibody of the invention by any of the methods described in detail above more than 1, more than 2, more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or more than 10 days following administration of the antibiotic agent. In one embodiment, the method may comprise administration of a first composition comprising a synthetic antibody of the invention by any of the methods described in detail above and a second composition comprising an antibiotic agent concurrently. In one embodiment, the method may comprise administration of a first composition comprising a synthetic antibody of the invention by any of the methods described in detail above and a second composition comprising an antibiotic agent concurrently. In one embodiment, the method may comprise administration of a single composition comprising a synthetic antibody of the invention and an antibiotic agent.

[0319] Non-limiting examples of antibiotics that can be used in combination with the synthetic antibody of the invention include aminoglycosides (e.g., gentamicin, amikacin, tobramycin), quinolones (e.g., ciprofloxacin, levofloxacin), cephalosporins (e.g., ceftazidime, cefepime, cefoperazone, cefpirome, ceftobiprole). antipseudomonal penicillins: carboxypenicillins (e.g., carbenicillin and ticarcillin) and ureidopenicillins (e.g., mezlocillin, azlocillin, and piperacillin), carbapenems (e.g., meropenem, imipenem, doripenem), polymyxins (e.g., polymyxin B and colistin) and monobactams (e.g., aztreonam).

[0320] The present invention has multiple aspects, illustrated by the following nonlimiting examples.

[0321] Examples

[0322] The present invention is further illustrated in the following Examples. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. Thus, various modifications of the invention in addition to those shown and described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

[0323] Example 1: IN VIVO DELIVERY AND PROTECTION BY ENGINEERED SYNTHETIC DNA-ENCODED INFLUENZA Hl HA HEAD-DIRECTED MONOCLONAL

[0324] ANTIBODIES IN MICE.

[0325] Monoclonal antibody (MAb) therapy is revolutionizing modem medicine, for cancer, and autoimmune treatment among others with more than 200 MAbs approved for clinical use or under review worldwide. For infectious disease, monoclonal antibodies received initial emergency use authorization during the COVID-19 pandemic and the recent licensure of nirsevimab for respiratory syncytial virus prophylaxis opens a new paradigm for mAh delivery against infectious diseases. Routine mAh delivery for seasonal infections like influenza A viruses could provide a significant public health benefit. However, the global potential of MAbs remains limited due to developability limitations, manufacturing costs, delivery route challenges, and the need for cold chain storage. In vivo gene delivery platforms, including synthetic plasmid DNA vectors can be engineered to encode antibody heavy' chain and light chain genes for direct delivery'. Synthetic DNA is simple to manufacture, temperature-stable, and DNA-encoded antibodies (DMAbs) are therefore a promising alternative modality for in vivo delivery of protective mAb.

[0326] Influenza viruses cause acute respiratory^ illnesses and are a significant cause of morbidity' and mortality' in all age groups, particularly young children and older adults, and influenza-related illnesses have returned to pre-COVID pandemic levels. As an approach to influenza prophylaxis, we engineered 11 different synthetic DMAbs encoding human IgGl monoclonal antibodies targeting multiple epitopes on the H1N1 influenza A virus hemagglutinin (HA) protein globular head. DMAbs were sequence optimized and then tested for their ability to express in vitro. Then, in vivo DMAb expression was evaluated following delivery of multiple doses (25-100pg) to BALB / c mice (n=5 / group), with peak serum levels reaching >20pg / mL at the highest dose. Serum binding and neutralization activity against A / HlNl / pdm2009 lineage viruses was detected, identifying 2 DMAbs with potent activity comparable to a previously published DMAb, DMAb-2-12C. These top 2 DMAbs were next administered to DBA / 2 mice (n=15 mice / group. 6.25-25pg doses) alone and in combination. achieving expression levels between l -5pg / mL by day 5 post-administration. The mice were challenged intranasally on Day 6 post-administration with 10LD50 of H1N 1 A / Califomia / 07 / 2009 X179A. Protection against weight loss and mortality was observed for all DMAb treatment groups, including at the lowest doses. Interestingly, the lungs histopathological analysis and immunohistochemistry (IHC) staining indicated dosedependent interstitial pneumonia and presence of influenza NP antigen. Lesions and positive IHC staining were only seen in negative control groups and low dose DMAb groups, with groups receiving higher doses of DMAb not showing any lesions.

[0327] Taken together, our data demonstrate that synthetic DNA Hl HA head-directed monoclonal antibodies are functional and offer protection in mice against lethal H1N1 pdm2009 infection, even at low doses. Our data are supportive towards translation of this approach to larger animal models with consideration for further translational development. DMAb delivery7offers important advantages for seasonal influenza prevention including long-term expression, potentially facilitating access to significant US. and global populations.

[0328] Example 2: DEVELOPING INFLUENZA H1HA HEAD-DIRECTED DNA-ENCODED MONOCLONAL ANTIBODIES AND EVALUATING THEIR PROTECTION IN AN Hl INFLUENZA CHALLENGE MODEL.

[0329] In recent years, monoclonal antibodies (mAbs) have been increasingly used in both the cancer and infectious disease spaces. 200 MAbs have been approved for clinical use or under review worldwide. Several monoclonal antibodies were approved for initial emergency use authorization and used as therapeutics during the SARS-CoV2 pandemic. Recently, a monoclonal antibody (Nirsevimab) targeting RSV was approved as a prophylactic for infants younger than 8 months of age.

[0330] Global access to mAbs is limited due to development hurdles, production limitations, stability, dosage, and costs. Expanding mAb access with gene-encoded antibodies addresses this problem by using gene delivery platforms which turn the body into a mAb producing "factory". DNA as a platform offers several advantages including local (ID and IM) delivery and local expression, transient expression (does not integrate), and temperature stable formulations / easy delivery7.

[0331] Influenza cases are back to pre-pandemic levels in some regions. Seasonal influenza includes A / H1N1, A / H3N2, B / Y amagata, and B / Victoria lineage viruses. In addition to updated yearly vaccines, additional approaches including protective monoclonal antibodies that can cover a flu season would be beneficial for many higher-risk groups.

[0332] DMAb-FISW086 has a comparable neutralization IC50 titre to dMAb-2-12C (Figure 1). Neutralization was performed on heat inactivated sera incubated with 100 TCID50 of H IN 1 / Bethesda (received from NIH). An overnight assay was performed according to the WHO influenza assay guidelines. Presence or absence of infection was detected by an anti-influenza NP ELISA. dMAb-FISW086 was evaluated using a DBA / 2 / CalO9 lethal model (Figure 2). These data demonstrated that dMAb-FISW086 is protective and prevents weight loss at very- low doses.

[0333] FISW086 dMAb dosage was evaluated (Figure 3). No treatment: interstitial pneumonia with dense cellular infiltrate, stains positive for IAV-NP. dMAb-FISW086 12.5 pg and 25 pg: mild interstitial pneumonia and stain negative for IAV-NP. dMAb-FISW086 6.25 pg and 3. 12: mild interstitial pneumonia and stains positive for IAV-NP around airways. Lung analysis enables a better visualization of dMAb effectiveness to support down-selection and additional development.

[0334] DMAb-LPAF-021 was evaluated in a DBA / 2 / CalO9 lethal model (Figure 4). These data demonstrate that dMAb-LPAF021 shows protection against weight loss and signs of disease at very- low doses. LPAF-021 is an additional mAb / dMAb that binds to a different epitope compared to FISW086.

[0335] LPAF021 dMAb dosage was evaluated (Figure 5). No treatment: interstitial pneumonia with dense cellular infiltrate, stains positive for IAV-NP. dMAb- LPAF021 25 pg: mild interstitial pneumonia and stains negative for IAV-NP. dMAb-LPAF021 12.5, 6.25pg and 3.12: moderate to severe interstitial pneumonia and stains positive for IAV-NP. Lung analysis enables a better visualization of dMAb effectiveness to support down-selection and additional development. dMAb cocktail delivery- was evaluated in the DBA / 2 lethal model (Figure 6). These data demonstrate that that a combination of the two dMAbs may improve protection synergistically at lower doses. LPAF-021 is an additional mAb / dMAb that binds to a different epitope compared to FISW086.

[0336] The combination dMAb dosage was evaluated (Figure 7). No treatment: interstitial pneumonia with dense cellular infiltrate, stains positive for IAV-NP. A combination of the 2 dMAbs protects against pneumonia at 25+25 pg and 12.5+12.5 pg and stain negative for NP. The 6.25+6.25 group shows mild pneumonia and stains positive for NP in small areas of the lung. The 3.12 + 3.12 group shows mild pneumonia in small areas of the lung and stains negative for flu NP.

[0337] These data demonstrate that these antibodies protect mice against lethal H1N1 pdm 2009 infection. Gene therapy platforms show potential for monoclonal antibody delivery as DNA encoded monoclonal antibodies targeting different pathogens have shown high expression levels over time and to be protective against infection. Thus, DNA-encoded monoclonal antibody cocktails have the potential to be used as a prophylactic against influenza disease.

[0338] Example 3: IN VIVO DELIVERY AND PROTECTION BY ENGINEERED SYNTHETIC DNA-ENCODED INFLUENZA A NEURAMINIDASE (NA)-DIRECTED MONOCLONAL ANTIBODIES IN MICE.

[0339] The data demonstrate that synthetic DNA neuraminidase (NA)-directed monoclonal antibodies are functional and offer protection in mice against A / Califomia / 07 / 2009 X179A infection, with better protection in the lungs (Figure 8). The data are supportive towards translation of this approach to larger animal models with consideration for further translational development. DMAb delivery' offers important advantages for seasonal influenza prevention including long-term expression, potentially facilitating access to significant US. and global populations.

[0340] Example 4: SYNTHETIC DNA CO-IMMUNIZATION WITH VACCINE-ALIGNED COMMON CONSENSUS NUCLEOPROTEIN AND HEMAGGLUTININ PROTECTS MICE ANGAINST LETHAL INFLUENZA INFECTION WITH A SINGLE IMMUNIZATION.

[0341] There is an urgent need for influenza vaccine strategies that enhance protection against influenza virus drift and across different subtypes. The conserved viral nucleoprotein (NP) is the most abundant viral protein and a target for broadly protective cellular immune responses. Guided by annual WHO-recommended seasonal vaccine strains, we engineered synthetic DNA vaccine candidates encoding vaccine-aligned common consensus (VACC) immunogens designed to represent immune diversity7of seasonal H1N1 and H3N2 virus NP proteins post-HlNlpdm09 and 2000-2022, respectively (pVACC-NPH1; pVACC-NPH3). Both NPH1and NPH3DNA vaccines induced robust cellular immune responses in mice, including the induction of durable responses. Immunization with a single dose of either DNA vaccine 14 days prior to lethal A(HlNl)pdmO9 virus challenge provided protection against mortality. Co-administration of pVACC-NPH3with an HA- expressing DNA vaccine (pHAH1) afforded improved protection against morbidity and mortality in a high-dose challenge model. These data highlight the potential of heterologous cellular immunity induced by engineered NP immunogens to complement HA- based approaches to significantly improve challenge outcomes.

[0342] Introduction

[0343] Seasonal influenza viruses infect approximately 1 billion people each year (Nair et al., Lancet (2011) 378(9807): 1917-30), causing respiratory' illnesses across both hemispheres. An estimated 3-5 million of these cases result in severe illness, with 290- 650,000 deaths annually (WHO. Influenza (Seasonal) - Fact Sheet: World Health Organization). Although vaccination is the safest and most effective means of influenza infection control, yearly genetic variation within the major influenza A virus (IAV) H1N1 and H3N2, and B virus strains necessitates annual reformulation. Broad and universal influenza vaccines are urgently needed to protect from circulating and newly emerging influenza viruses. In addition to strategies that induce broadly protective antibodies against the viral surface hemagglutinin (HA) protein, synthetic immunogen approaches that direct protective immunity to target highly conserved epitopes or proteins could provide important adjunctive protection to decrease pathogenesis and severe disease.

[0344] The influenza nucleoprotein (NP) is the most abundant internal protein and the major component of the virion ribonucleoprotein complex. It plays a critical role in viral replication, involving organization of RNA packing, nuclear trafficking, vRNA transcription, and replication (Hu et al., Current topics in medicinal chemistry' (2017) 17(20):2271-85). NP is well-conserved within influenza subtypes, making it a promising target for inducing cellular immune responses. NP has been shown to induce robust CD8+ T cell responses in preclinical models (Y ewdell et al., Proceedings of the National Academy of Sciences (1985) 82(6): 1785-9; Zhou et al., Molecular Therapy (2010) 18(12):2182-8) and humans (McMichael et al., Journal of general virology (1986) 67(4): 719-26). Computational modeling of influenza isolates has revealed stretches of highly conserved amino acids within NP and peptide vaccines based on such epitopes elicit robust CD8+ T cell responses and are protective against IAV challenge in mice (McGee et al.. Journal of medical virology (2022) 94(6):2578-87). Epidemiological studies indicate that anti-NP CD8+ T cell immunity can contribute to protection from severe disease in humans (Epstein, The Journal of infectious diseases (2006) 193(l):49-53). These data suggest that NP based therapies have the potential to elicit broad anti-influenza cellular immunity'.

[0345] Synthetic plasmid DNA vaccines have advanced significantly over the past ten years, demonstrating robust induction of humoral and cellular immune responses (Gary and Weiner, Current Opinion in Immunology (2020) 65:21-7). The first DNA vaccine received EUA for use in humans during COVID-19 (Khobragade et al., The Lancet (2022) 399(10332): 1313-21) and several T cell-based DNA vaccines are being evaluated for infectious diseases and delivery of cancer neoepitopes (Yarchoan et al.. Nature medicine (2024): 1-10.) to elicit CD8+ T cell responses. Current inactivated vaccines elicit poor CD8+ T cell responses compared to live attenuated influenza vaccines (LAIV) (Blazevic et al., Human immunology (2000) 61(9):845-9 ; Hoft et al., Journal of Infectious Diseases (2011) 204(6):845-53; Cox et al., Scandinavian journal of immunology’ (2004) 59(1): 1-15). Although LAIV vaccines can induce CD8+ T cell responses, the master donor virus used to make all LAIVs contains the internal genes, including NP, of A / Ann Arbor / 6 / 60 or A / Leningrad / 17 / 57 H2N2 viruses and is thus mismatched to modem circulating strains. To this end, studies matching LAIV vaccines to currently circulating viruses can increase induction of CD8+ T cell responses (Smith et al., J Virol (2020) 94(4)). Building on this prior research, we hypothesized that plasmid DNA-encoded NP consensus sequences can expand breadth of protection, eliciting broad cellular immune responses that can reduce IAV pathogenesis.

[0346] Here, the design and evaluation of synthetic IAV-NP immunogens engineered based on WHO-recommended vaccine strains to induce robust anti -influenza cellular immunity in vivo are described and tested. Tyvo DNA-encoded vaccine-aligned common consensus (VACC) immunogens representing the NPs from seasonal A / H1N1 (VACC-NPH1) or A / H3N2 (VACC-NPH3) viruses induced robust cellular immune responses, with both independently providing single dose protection against mortality in mice intranasally challenged with an A(H1N1) pdm09 vims. Heterologous pVACC- NPH3combination with plasmid-encoded hemagglutinin from H IN 1 A / Califomia / 07 / 2009 (pHAH1) afforded complete protection from lAV-associated morbidity and mortality’, further highlighting the potential for synthetic VACC-NPXcandidates to reduce pathogenesis and provide immune protective benefit across IAV subtypes.

[0347] Methods

[0348] Plasmid design: The amino acid sequences for NP proteins from WHO recommended H1N1 and H3N2 vaccine strains (World Health Organization (2025) [cited 2025 March 24, 2025], Available from: https: / / www.who.int / tearns / global-influenza- programme / vaccines / who-recommendations) were downloaded from the GISAID.org database. H1N1 NP accession #: A / New Caledonia / 20 / 1999 (EPI ISL 649), A / Solomon Islands / 3 / 2006 (EPI224787), A / Brisbane / 59 / 2007 (EPI ISL 154495), A / Califomia / 7 / 2009 (EPI ISL 391380), A / Michigan / 45 / 2015 (EPI ISL 199532), A / Brisbane / 02 / 2018 (EPI ISL 344858), A / Wisconsin / 588 / 2019 (EPI ISL 404527), A / Hawah / 70 / 2019 (EPI ISL 397028). H3N2 NP accession #: A / Moscow / 10 / 99 (EPI ISL 2695), A / Fujian / 411 / 2002 (EPI ISL 107711), A / Califomia / 7 / 2004 (EPI ISL 113070), A / Wisconsin / 67 / 2005 (EPI ISL 154528), A / Brisbane / 10 / 2007 (EPI ISL 176458), A / Perth / 16 / 2009 (EPI ISL 176456), A / Victoria / 361 / 2011 (EPI ISL 101506). A / Switzerland / 9715293 / 2013 (EPI ISL 166310), A / Hong Kong / 4801 / 2014 (EPI ISL 233740), A / Singapore / INFIMH- 16-0019 / 2016 (EPI2397166), A / Kansas / 14 / 2017 (EPI ISL 292575), A / Hong Kong / 45 / 2019 (EPI ISL 347938). H1NP or H3NP vaccine-consensus designs were constructed through sequence alignment analysis in MEGA 11.0. 10 (Tamura et al., Mol Biol Evol (2021) 38(7):3022-7) using ClustalW alignment and an unrooted phylogenetic tree was generated using the maximum-likelihood method, with maximum parsimony (Patel et al., J Infect Dis (2019) 219(4):544-55). Pairwise distances were calculated in MEGA. 11.0.10. Sequence identity visualization was performed in Treeviewer (Bianchini et al., Ecol Evol (2024) 14(2):el0873). Additional alignment of sequences were performed in Geneious Prime (version 2023.2.1).

[0349] Cell lines and virus propagation: Influenza A Virus, A / Califomia / 07 / 2009 NYMC X-179A (H1N1) pdm09, FR-246, was obtained through the International Reagent Resource, Influenza Division, WHO Collaborating Center for Surveillance, Epidemiology and Control of Influenza, Centers for Disease Control and Prevention, Atlanta, GA, USA. MDCK-SIAT1 cells (Sigma Cat# 5071502) were maintained in MEM Eagle’s with 1% Pen / Strep, and 2% FBS. For virus propagation, cell monolayers were infected with MOI 0.001 of A / Califomia / 07 / 2009 X179A in the presence of 2 pg / mL TPCK-treated Trypsin (ThermoFisher Cat# 20233) and maintained with 1% Pen / Strep, 0.3% BSA for 3 days. Virus was collected and ultracentrifuged on a sucrose gradient to prepare mouse challenge stocks. Challenge stocks were titered on MDCK-SIAT1 cells and an initial mouse LD50 experiment was performed to determine the minimum infectious dose.

[0350] Animals, Immunization, and Challenge: C57BL / 6J and DBA / 2 mice were purchased from the Jackson Laboratory and were housed in the Wistar Institute Animal Facility . All procedures were done in accordance with the guidelines from the Wistar Institute Animal Care and Use Commitee. Between 2 pg to 10 pg of DNA plasmid encoding the VACC-NPH1or VACC-NPH3or a full length HA DNA (HAH1, A / Califomia / 07 / 2009 (Tursi et al., Cell Rep Med (2025): 102035)) with or without a DNA plasmid encoding for the molecular adjuvant IL12, in 30 pL water was injected in the tibialis anterior (TA) muscle, followed by delivery of two 0.1 Amp electric constant current square-wave pulses by the CELECTRA-3P electroporation device (Inovio Pharmaceuticals) to increase transfection efficiency. Immunized or naive DBA / 2 mice were intranasally infected with 10LD50 or 100LD50 of A / Califomia / 07 / 2009 X179A respectively 50pl minimum essential medium (MEM) Eagle’s (without antibiotics). Mice were then monitored for the subsequent 21 days, for weight loss and mortality. Any mouse reaching 80% of their original body weight was considered to have reached humane endpoint and was subsequently euthanized. A subset of mice (n=3 per group) was euthanized on day 6 post infection, and lungs were collected for histopathological analysis. The vaccine and challenge schedules are indicated in each figure.

[0351] Western Blot: HEK293T cells were cultured in DMEM medium with 10% FBS at 37°C / 5% CO2 condition and transfected with pDNA using Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific Cat# L300000) following the manufacturer’s protocol. Forty-eight hours later, supernatant and cell lysates were harvested using lx cell lysis buffer (Cell signaling Cat# 9803). Proteins were separated on a 4-12% BIS-TRIS gel (Thermo Fisher Scientific Cat# NP0322BOX), then following transfer, blots were incubated with an anti-NP monoclonal antibody (Thermo Fisher Cat# PA5-32242), then visualized with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Sigma Cat# SAB3701359).

[0352] Flow Cytometry': Immunized mice were euthanized, and spleens and lungs were harvested and stored in RPMI 1640 media (Invitrogen Cat# 11875093). Spleens were processed to single-cell suspension and red blood cells were removed by ACK lysing buffer (Gibco Cat# A1049201). Lungs were processed using the lung dissociation kit / GentleMACS system (Miltenyi Cat# 130-095-927) according to manufacturer’s instruction, red blood cells were removed by ACK lysing buffer (Gibco Cat# A1049201), and single cell suspensions were isolated. Cells were then filtered and counted before being plated for flow cytometry. Cells (1,000,000 per well) were seeded in 100 pL of RPMI 1640 supplemented with 10% FBS and 1% Penicillin / Streptomycin (RIO). Cells were stimulated with peptides spanning the NPH1, the NPH3, or an H1N1 HA (A / Califomia / 07 / 2009) at a final concentration of 5 pg / mL per peptide in the presence of Protein Transport Inhibitor (eBioscience, San Diego, CA, USA Cat# 00-4980-03). Cell Stimulation Cocktail (eBioscience, San Diego, CA. USA Cat# 00- 4970-93) in R10 were used as negative and positive controls, respectively. Plates were incubated for 6 h at 37°C with 5% CO2. Splenocytes were stimulated by peptides for 5 h with protein transport inhibitor (eBioscience, San Diego, CA, USA Cat# 00-4980-03). After stimulation, cells were stained with LIVE / DEAD zombie aqua for viability. CD3, CD4, CD8, TNF-a. IFN-y, and IL-2 fluorochrome conjugated antibodies (all from BioLegend) were used for surface and intracellular staining. The samples were run on aBD FACSymphony™ A5 SE flow cytometer (BD Biosciences) and analyzed by FlowJo software. Gates were set using FMOs for each stain. Data were exported and analyzed in GraphPad Prism 10.

[0353] ELISpot: Isolates splenocytes and pulmocytes were subjected to IFNy ELISpot assay according to the manufacturer’s instructions (Mabtech). Briefly, plates were washed four times with sterile PBS and blocked with R10 media for two hours. Wells were seeded in duplicate with 200,000 cells suspended in lOOpL R10. Cells were stimulated with 5pg / ml of either peptide pools representing the entire protein for either NPH1, NPH3or HAH1. Negative and positive controls were stimulated with DMSO or phorbol myristate acetate / ionomycin (PMA / Iono) respectively. Plates were incubated at 37°C in 5% CO2 for 18 hours and were then developed following the manufacturer’s protocol. Plates were scanned and counted using the Mabtech IRISTM FluoroSpot / ELISpot reader.

[0354] Histopathology and Immunohistochemistry’: Whole murine lungs were collected into 10% buffered neutral -buffered formalin for routine histopathological processing. Formalin fixed tissues were paraffin embedded and 4 pm sections were cut and routinely stained with Hematoxylin and Eosin. Immunohistochemical detection was performed on 4 pm tissue sections using an anti-IAVNP antibody (Thermo Fisher Cat# PA5- 32242). Whole slides were scanned using a Hamamatsu Nanozoomer S60 slide scanner and analyzed using NDP.view 2. Scale bars equal 2.5 mm on whole slide lung images and 50 pm on lung section images.

[0355] RNA-seq: Lung tissue samples from murine paraffin-embedded blocks were used for RNA extraction, with integrity confirmed using the Agilent Bioanalyzer (RIN > 7). Libraries were prepared using the QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen Cat# 191.24) and sequenced on an Illumina platform. Sequencing reads were preprocessed using a custom Python script to trim low-quality bases and adapters, followed by alignment to the mmlO genome using Bowtie2 within the RSEM pipeline (vl.3.3). Only reads mapping to coding regions were retained. Raw counts and TPM values were generated for downstream analyses. Differential gene expression analysis was conducted using DESeq2 (vl.38.0). Genes with fewer than 10 raw counts were excluded, and DEGs were identified using FDR < 5% and |log2 fold change] > 3. Functional enrichment was performed using Gene Ontology, KEGG pathways, and Ingenuity Pathway Analysis (IP A). Computational analyses were conducted on a Linux-based high-performance computing environment with tools including Python scripts, Bowtie2 (v2.4.5), RSEM (vl.3.3), DESeq2 (vl.38.0), and IP A.

[0356] Software and statistical analysis: Data was represented in GraphPad Prism version 10. All sequence alignments were determined in MEGA 11.0. 10 (Tamura et al., Mol Biol Evol (2021) 38(7):3022-7) and Treeviewer (Bianchini et al., Ecol Evol (2024) 14(2):el0873), flow cytometry data was analyzed using FlowJo version 10.10.0. Image slides were scanned using a Hamamatsu Nanozoomer S60 slide scanner and analyzed using NDP.view vs2. Details on statistical analysis are included in the legend for each figure.

[0357] Results

[0358] Design and expression of VACC-NPXimmunogens: NP amino acid sequences for annual seasonal A / H1N1 (NPH1, Figure 9A) and A / H3N2 (NPH3, Figure 9D) vaccines strains were obtained from GISAID.org and aligned to produce unrooted phylogenetic trees (Figures 9B and 9E, strains listed in Figure 16, alignments in Figure 17 and Figure 18). Tw o separate vaccine-aligned consensus construct (VACC) designs were generated, with weighting of amino acids towards more recent NPs. The overall pairwise distances w ere determined to be <0.2% for post-HlNlpdm09 IAV-NPH1s and <1.1% for IAV-NPH3s. Sequences were codon-optimized for mammalian expression and subcloned into thepVAXl plasmid DNA backbone to generate pVACC-NPHl and pVACC-NPH3 constructs (Figures 9C and 9F). mRNA expression of VACC-NPXwas confirmed via quantitative PCR (Figure 1G) following in vitro transfection. Protein expression in transfected cells was confirmed via supernatant western blot (Figure 9H), and immunofluorescence staining of IAV-NP (Figure 91). Together these data demonstrate that the consensus alignment approach generates novel synthetic molecules that express in vitro and are detected by commercial anti-NP antibodies. A single immunization with DNA-encoded pVACC-NPxvaccines induces strong cellular responses and supports protection from influenza-associated morbidity and mortality in vivo: C57BL / 6 mice were immunized once with 10 pg of pVACC-NPH1or pVACC-NPH3immunogens and cellular responses were evaluated by ELISpot assay fourteen days later (Figure 10A). pVACC- NPH1induced significant H1N1 NP-specific IFNy spotforming units (SFU) in the spleens of immunized mice as compared to empty plasmid (pVaxl) immunized controls (Figure 10B). Similarly, pVACC-NPH3resulted in significant induction of H3N2 NP-specific IFNy responses (Figure 10C). These data demonstrate that pVACC-NPxconstructs induce strong cellular immunity7in vivo.

[0359] To evaluate the protective efficacy of these constructs, DBA / 2 mice were immunized once with 10 pg of pVACC-NPH1, pVACC-NPH3, or left unimmunized (naive), and challenged fourteen days later with 10 LD50 of H1N1 A / Califomia / 07 / 2009 / X179A (Figure 10D). 90% survival was observed among receiving pVACC-NPH1and 100% survival among pVACC-NPH3immunized animals while all naive animals succumbed to infection (Figure 10E). All pVACC-NPximmunized animals displayed significant weight loss (Figure 10F) and this was reflected by hematoxylin & eosin (H&E) staining of lungs harvested 6 days post-infection (Figure 10G). Dense cellular infiltrates were observed in the lungs of naive and pVACC-NPH1immunized animals. pVACC-NPH?immunized animals displayed decreased cellular infiltrates and increased airway space (Figure 10G). Similarly, when sections were stained for H1N1 NP antigen, naive animals displayed significant NP-positive staining throughout their lungs. pVACC-NPH1immunized animals had decreased NP antigen and only minimal staining was observed in the lungs of pVACC-NPH3immunized mice (Figure 10H). Together, these data highlight the potential for a VACC-NPXimmunogen to provide benefit against disease and death within 2 weeks of a single immunization.

[0360] Epitope mapping of VACC-NPx-induced cellular responses: Epitope mapping was performed with a matrix of overlapping peptide pools to identify immunodominant peptides by ELISpot for NPH1and NPH3(Figure 18) in mice co-immunized with pIL-12. We identified the linear peptides ASNENVETM among NPH1peptides (Figure 18B and Figure 18C) and ASNENMDNM among NPH3 peptides (Figure 18D and Figure 18E), consistent with those described for murine H2-Db in the literature. pVACC-NPH3also elicited strong responses to the SAAFEDLRLLSFIRG peptide reported by Lambe et al. (Lambe et al., Sci Rep (2013) 3: 1443). These data demonstrate that the pVACC-NPxconstructs can elicit responses consistent with previously identified epitopes, as well as can expand unique responses. pVACC-NPxantigens are amenable to co-deli\ ery with HA immunogens: Current seasonal influenza vaccines are either inactivated virus, live attenuated virus, or recombinant protein vaccines, all of which contain large quantities of HA antigen. As both pVACC-NPH1and pVACC-NPH3induced similar immunogenicity and protective efficacy, we selected the heterologous pVACC-NPH3for evaluation alone and in combination with a plasmid-encoded H1N1 HA antigen (pHAH1). C57BL / 6 mice were immunized once with either 10 pg pVACC-NPH3, 2 pg pHAH1, 10 pg of pVACC-NPH3plus 2 pg of pHAH1, or both immunogens and 0.5 pg of pIL-12 (Combo), and cellular responses were quantified fourteen days later (Figure 11 A). Splenocytes stimulated with NP peptides induced robust IFNy responses in the spleens of pVACC-NPH3and combo immunized mice (Figure 11B). Mice administered empty plasmid or pHAH1alone did not display significant T cell responses (Figure 11B). When splenocytes were stimulated with matched H1N1 HA peptides, only the pHAH1and combo-immunized mice induced significant SFUs (Figure 11C, p<0.0001 and p<0.01 , respectively). These data demonstrate that co-delivery of pVACC-NPH3and pHA can elicit cellular responses against both antigens simultaneously.

[0361] Cellular immune responses induced by this combination were profiled approach by intracellular cytokine staining (ICS). Splenocytes stimulated with NP peptides induced significant increases in the frequency of IFNy+ effector CD8+ T cells in splenocytes from mice receiving pVACC-NPH3alone, pVACC-NPH3plus IL-12, pVACC-NPH3plus pHAH1, or the combination of antigens and pIL-12 but not in those from pVaxl or pHAH1- only immunized mice (Figure 1 IB). There was also a significant increase in CD107a+IFNy+ effector CD8+ T cells among these mice (Figure 1 IB). Conversely, when splenocytes were stimulated with matched HA peptides, significant frequencies of IFNy+ (Figure 11C) and CD107a+IFNy+ (Figure 11C) effector CD8+ T cells were observed only in the splenocytes of mice immunized with pHAH1alone, co-immunized with either pHAH1and pVACC-NPH3, or co-immunized with the combination of pHAH1. pVACC-NPH3. and pIL-12.

[0362] Cellular responses were assayed from lungs as it is the primary site of influenza infection and replication. Lung lymphocytes (pulmocytes) stimulated with NP peptides induced significant increases in the frequency of IFNy+ effector T CD8+ T cells in mice receiving pVACC-NPH3alone. pVACC-NPH3plus IL-12, pVACC-NPH3plus pHAH1. or the combination of antigens and pIL-12 but not in those from pVaxl or pHAH1-only immunized mice (Figure 1 IB). There was also a significant increase in CD107a+IFNy+ effector CD8+ T cells among these mice (Figure 1 IB). Conversely, when pulmocytes were stimulated with matched HA peptides, we observed significant frequencies of IFNy+ (Figure 11C) and CD107a+IFNy+ (Figure 11C) effector CD8+ T cells only in the pulmocytes of mice immunized with pHAH1alone, co-immunized with either pHAH1and pVACC-NP113. or co-immunized with the combination of pHAH1, pVACC-NPH3, and pIL-12. While HA- specific responses were attenuated when pVACC-NPH3was co-delivered with pHAH1, HA- specific responses continued to be detectable among animals receiving the Combo vaccination regimen. These data suggest that combination delivery of pVACC-NPxantigens with HA antigens can elicit robust cellular responses in both the periphery and mucosa.

[0363] Heterologous pVACC-NPH3enhances protective efficacy of the pHAH1DNA vaccine against IAV A(HlNl)pdmO9 challenge: The abil ity of a synthetic VACC-NPXimmunogen to provide adjunctive protection when administered in combination with an HA vaccine was tested. In the DBA / 2 mouse model, a single immunization with pHAH1alone induces complete protection against morbidity and mortality from a 10LD50 homologous A / Califomia / 07 / 2009 X179A challenge. At the higher challenge inoculum of 100LD50 mice immunized with 10 pg pHAH1are partially protected (70%) from death (Figure 18A and Figure 18B) but observed substantial weight loss post-challenge (Figure 18C). This sub- protective model was next used to evaluate the protective efficacy following combination delivery of the pVACC-NPH3and pHAH1vaccines.

[0364] DBA / 2 mice were immunized once with pHAH1alone (10 pg), pVACC-NPH3alone (10 pg), or co-immunized with pHAH1and pVACC-NPH3(10 pg each). All immunization groups also received a standard dose of pIL-12 (0.5 pg) (Figure 12A). Animals were challenged fourteen days post-immunization with 100LD50 A / Cahfomia / 07 / 2009 X179A. Animals which received pVACC-NPH3alone succumbed to this lethal challenge by day 7, as did naive animals, however 100% of mice which received pHAH1alone or the combination vaccine survived challenge (Figure 12B). All pHAH1immunized animals lost significant weight, but survived challenge (100%). Interestingly, only the Combo group afforded complete protection from both weight loss (Figure 12B) and mortality (Figure 12C). H&E staining revealed dense cellular infiltrates in the lungs of naive and pVACC-NPH3-only immunized mice (Figure 12D). pHAH1-only and combination-immunized mouse lungs displayed more open airway space but had intermediate cellular infiltration and modest evidence of alveolar wall thickening (Figure 12D). When sections were stained for NP antigen, naive animals had significant, dispersed NP antigen staining (Figure 12E). pVACC- NPH3- only immunized mouse lungs exhibited dense NP antigen staining which was localized to the alveolar spaces. In pHAH1-only immunized mouse lungs, NP staining was faint and dispersed, whereas Combination-immunized lungs display minimal NP positivity (Figure 12E). Taken together, these data support that the combination delivery of NP with HA antigens can improve challenge outcome.

[0365] Lung gene expression during infection following administration of pHAH1and pVACC-NPH3DNA vaccines. Differential gene expression between untreated and DNA vaccine treated mice was profiled post-infection using Quantseq on RNA extracted from lung tissue scrolls of mice immunized with the pHAH1and pVACC-NPH3DNA vaccines 6 days post-infection with 10 LD50 A / Califomia / 07 / 2009 X179A (Figure 13A). PCA and gene analysis show distinct clustering of the two immunized groups (pHAHl and pVACC-NPH3) differently from infected non- immunized mice (Figure 13B and Figure 13C), Volcano plots show a total of 2088 downregulated and 1694 upregulated genes and 2716 downregulated and 2301 upregulated genes in the pHAH1and pVACC-NPH3groups respectively (Figure 13D and Figure 13E). Ingenuity pathway analysis (IP A) indicated the activation or inhibition of several pathways related to viral infection, immune cells, cytokines and others in animals immunized with the pHAH1or pVACC-NPH3vaccines as compared to non-immunized (infected) mice (Figure 13F and Figure 13G). Taken together, these data highlight the ability of both DNA vaccines to independently induce responses that lead to antiviral control during infection.

[0366] DNA-encoded VACC-NPXantigens induce durable T cell responses in mice: C57BL / 6 mice received two immunizations separated by three weeks (10 pg per antigen) to evaluate cellular immune responses induced by pVACC-NPxcandidates (Figure 14A and Figure 14F). DNA antigen co-formulation with plasmid-encoded IL-12 (pIL-12) to enhance cellular (Kalams et al., The Journal of infectious diseases (2013) 208(5):818-29; Vonderheide et al.. Journal for ImmunoTherapy of Cancer (2021) 9(7)) and humoral (De Rosa et al., JCI insight (2020) 5(13)) responses in humans and in preclinical models (Sin et al.. The Journal of Immunology (1999) 162(5):2912-21 ; Hirao et al.. Vaccine (2008) 26(25):31 12-20; Boyer et al., Journal of medical primatology (2005) 34(5-6):262-70; Chattergoon et al., Vaccine (2004) 22(13-14): 1744-50)) has been previously reported. Therefore, parallel groups of mice were co-immunized with pIL-12 to study ability to adjuvant NP-directed cellular responses. T cell responses were detected by IFNy ELI SPOT assay with the spleens and lungs of mice following immunization. Mice receiving pVACC-NPH1demonstrated increased IFNy responses in the spleen compared to control animals with a further significant increase in responses when combined with pIL-12 (Figure 14C). Robust T cell responses were detected from lung pulmocytes in both vaccinated groups compared to control (Figure 14D). Both pVACC-NP™ DNA vaccine groups demonstrated significantly increased NP-directed cellular immune responses over control animals, with pIL-12 further enhancing IFNy responses in cells assayed from both tissues (Figure 14E and Figure 14F).

[0367] To evaluate the longevity of pVACC-NPx-induced cellular responses, mice receiving the same regimen were rested for -200 days (6 months) (Figure 14B). At this memory timepoint, significant anti-NP responses in the spleen and lungs for both pVACC- NPH1(Figures 14G and Figure 14H, respectively) and pVACC-NPH3(Figure 141 and Figure 14J, respectively) compared to naive controls. Co-delivery of pIL-12 demonstrated long-term enhancement of IFNy secretion in both compartments compared with animals receiving pVACC-NPH1(Figure 14G and Figure 14H, respectively). Significantly higher T cell responses were detected in animals receiving pVACC-NPH3and pIL-12 (Figure 14J. p<0.01) These data indicate that VACC-NPXDNA vaccines can elicit robust and long-lived cellular responses in vivo and highlight the potent contribution of molecular adjuvant pIL-12 to enhancing immunity directed against IAV-NP.

[0368] Discussion

[0369] The conserved IAV nucleoprotein is an attractive target for broad and universal influenza strategies. In humans, anti-NP cytotoxic T lymphocyte responses can reduce pathogenesis and confer important heterosubtypic protection (Amoah et al., J Virol (2024) 98(8):e0071124). Conventional seasonal influenza vaccines induce antibodies primarily directed towards the surface HA glycoprotein and HAI titers >1:40 are associated with protection in 50% of people. This protection is most optimal against matched and minimally mutated strains. However, yearly antigenic drift can significantly impact humoral antibody responses and multivalent combination with a of a potent CTL-targeting immunogen like NP could potentially address yearly diversity' by limiting infection- associated pathogenesis. Taking this into consideration, the design and immunogenicity of two new synthetic NP immunogens guided by the genetic sequences obtained from the seasonal IAV -H INI and IAV-H3N2 vaccine strains recommended by the WHO are described. This vaccine-aligned common consensus approach, or VACC, highlights the high conservation between NP proteins from IAV-H3 viruses over 20+ years and IAV-H1 strains post-pdm09. Synthetic DNA vaccination with these de novo immunogens induced strong T cell responses in the spleen and lungs and protected against lethal A / Califomia / 07 / 2009 X179A IAV-H1N1 challenge in mice. Co- delivery7of a DNA-encoded synthetic HAH1immunogen with the heterologous VACC-NP11’ immunogen is demonstrated to improve protection from weight loss and provide rapid protection in a highly stringent infection model.

[0370] Synthetic NP immunogens have been evaluated in various platforms including nucleic acids (DNA, mRNA), and viral vector-based platforms (adenovirus, MV A, and others). In addition to strain-matched designs, approaches to targeting IAV-NP include CD8+ T cell epitope-based strains and oligomerized forms (Leroux-Roels et al.. Lancet Infect Dis (2023) 23(12): 1360-9, Del Campo et al., Front Immunol (2021) 12:678483). Here, a synthetic consensus immunogen was designed with the goal of inducing broad protective responses. The number of sequenced circulating influenza strains has dramatically increased with advancements in sequencing technologies. One approach is to computationally align thousands of IAV NP sequences to generate single sequence (ref Xie et al below). For more the more variable surface HA and NA glycoproteins, this can be further stepwise divided into development of microconsensus sequences for HA and NA antigen designed to provide broad coverage within related phylogenetic branch points (Yan et al., Vaccine (2018) 36(22):3079- 89, Elliott et al., Hum Gene Ther (2018) 29(9): 1044-55). Comparatively, the overall diversity of IAV-NP within currently circulating human Hl and H3 subtypes is minimal, with high heterosubtypic conservation compared to HA and NA.

[0371] Since the 1970s, the WHO provides recommendations for the composition of seasonal influenza vaccines. This requires yearly surveillance involving analysis of clinical specimens, disease burden, and epidemiological data to understand representative viruses in the human population and their distribution by country7and region (Process of Influenza Vaccine Virus Selection and Development. Available from: https: / / apps.who.int / gb / pip / pdf files / Fluvaccvirusselection.pdf). The selected vaccine viruses could therefore be considered as representative of the diversity of major influenza viruses circulating in the human population in a current year. Using this as a guide, the VACC-NPXcandidates therefore encompass yearly NP variation. As highlighted, the current circulating human IAV-H3 viruses have varied minimally over the past 20 years (<1. 1%). In 2009, the introduction of the triple reassortant HlNlpdm2009 swine flu viruses into the human population resulted introduction of a classical swine H1N1 NP into humans, a significant antigenic shift (>10%). Therefore the VACC-NPH1design was focused based on post- HlNlpdm2009 viruses. To further address major antigenic shift events like in the case of HlNlpdm2009, additional consideration of animal (for example swine) Hl and H3 circulating strains would be valuable. Although there is no global body selecting vaccine strains for animals, similar surveillance of strains circulating in animals is being undertaken by agencies like the CDC, USDA, European CDC, WHO, WAOH, CEIRS program and others, alerting to emerging influenza strains with potential for zoonotic crossover into humans. Yearly monitoring and selection of predominantly circulating animal I AV would be valuable for narrowing down and selecting strains for inclusion in immunogen design. Such animal I AV -Hl and IAV-H3 NP immunogens could be incorporated as multivalent combinations to elicit broader cellular immune responses against potential emerging viruses. Interestingly, even with >10% distance, pVACC-NPH3affords comparable protection against a lethal H1N1 infection as pVACC-NPH1, highlighting the potential for our VACC design strategy across IAV subtypes with potential to temper major antigenic events.

[0372] These data show the protective potency of targeting the IAV-NP, achieving single dose protection 14 days following delivery in mice. Both pVACC-NPH1and pVACC- NPH?elicited robust T cell responses that were durable in mice. While NP generates a robust CTL response and can potentially contribute to humoral immunity (Krammer et al., Nature Reviews Immunology (2019) 19(6): 383-9), it remains likely that an HA immunogen component will be essential in IAV vaccine formulations to provide robust antibody- mediated protection. Combination of pVACC-NPH?with a plasmid-expressed H1N1 HA, robust T cell responses were observed against both HA and NP peptides. The pHAH1antigen alone induced complete protection in our low-dose IAV challenge model, however at higher challenge inoculums, pHAH1did not protect against weight loss and lung pathogenesis. When mice were co-immunized with pHAH1in combination with pVACC-NP11' and challenged in this high dose model, neither antigen alone provided complete protection. However, the coimmunized animals were completely protected from morbidity and mortality. These data indicate that pVACC-NPH3, and indeed other NP immunogens, can play a role to complement HA-based vaccine-induced immunity. Additional studies dissecting this synergy, likely due to T cell immunity would be interesting. Our approach generating synthetically designed NP antigens based on yearly vaccine stains can be broadly applied to other highly conserved influenza internal proteins with potential to generate strong CTL responses. Further study of the protective benefits of NP antigen co-delivery with seasonal HA- based vaccine regimens will be insightful. These together support the incorporation of the VACC design approach for continued development of broad and efficacious influenza interventions. Sequences

[0373]

[0374] SEQ ID NO:1 (NA) DMAb-2-12C (single plasmid)

[0375] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0376] AGTCCAGCTGGTCCAGTCAGGGGCCGAGGTGAAGAAGCCTGGCGAGTCCCTGAAGATCT

[0377] CTTGCAAGGGCAGCGGCAACGGCTTCACCACATACTGGATCGGATGGGTGCGCCAGGTG

[0378] CCAGGCAAGGGCCTGGAGTGGATGGGCATCATCTACCCAGACGATTCCGACACCAGGTA

[0379] TTCTCCCAGCTTTCAGGGCCAGGTGTCCATCTCTGCCGATAAGAGCATCTCCACCGCCTTC

[0380] CTGCAGTGGAGCTCCCTGAAGGCCAGCGACACAGCCATGTATTTTTGCGCAAGGCTGGG

[0381] CGACGTGGAGACAGCAATGGTGGGCCAGGATGCCTTCCACATCTGGGGCCAGGGCACCA

[0382] TGGTGACAGTGTCTAGCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCT

[0383] CTAAGTCTACCAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTG

[0384] AGCCAGTGACCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAG

[0385] CCGTGCTGCAGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTA

[0386] GCCTGGGCACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTG

[0387] GACAAGAAGGTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGC

[0388] ACCTGAGCTGCTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACT

[0389] GATGATCTCCCGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCC

[0390] CGAGGTGAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGC

[0391] CCAGGGAGGAGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCAC

[0392] CAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGC

[0393] CCCCATCGAGAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTAC

[0394] ACCCTGCCTCCAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGT

[0395] GAAGGGCTTCTATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGA

[0396] ACAATTACAAGACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCA

[0397] AGCTGACCGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGC

[0398] ACGAGGCCCTGCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGAGG

[0399] GGCAGAAAGAGGAGAAGCGGCTCCGGAGCAACCAACTTCTCCCTGCTGAAGCAGGCAG

[0400] GCGACGTGGAGGAGAATCCTGGACCAATGGTGCTGCAGACCCAGGTGTTCATCAGCCTG

[0401] CTGCTGTGGATCTCCGGCGCCTACGGCGAGATCGTGCTGACCCAGTCCCCAGGCACACTG

[0402] TCTCTGAGCCCTGGCGATCGGGCCACCCTGTCCTGTAGGGCATCCCAGAGCGTGAGCAGC

[0403] AGCTTCCTGGCCTGGTACCAGCAGAAGCCTGGCCAGGCCCCAAGGCTGCTGATGTATGG

[0404] AGCCTCCCGGAGGGCAACAGGCATCCCCGACAGATTCAGCGGCTCCGGCTCTGGAACCG

[0405] ACTTCACCCTGACAATCAGCCGGCTGGAGCCCGAGGACTTTGCCGTGTACTATTGCCAGC

[0406] AGTACGATTCCTCTCCCTTCACCTTCGGCGGCGGCACAAAGGTGGAGATCAAGAGAACA

[0407] GTGGCCGCCCCCAGCGTGTTCATCTTTCCACCCAGCGACGAGCAGCTGAAGTCCGGCACC

[0408] GCCTCTGTGGTGTGCCTGCTGAACAATTTCTACCCTCGGGAGGCCAAGGTGCAGTGGAAG

[0409] GTGGATAACGCCCTGCAGTCCGGCAATTCTCAGGAGAGCGTGACCGAGCAGGACTCCAA GGATTCTACATATAGCCTGAGCTCCACCCTGACACTGAGCAAGGCCGACTACGAGAAGC ACAAGGTGTATGCCTGTGAGGTCACCCACCAGGGGCTGTCAAGTCCAGTCACTAAAAGT TTCAATAGGGGAGAATGTTGA

[0410] SEQ ID NO:2 (AA) DMAB-2-12C (single plasmid)

[0411] MDWTWRILFLVAAATGTHAEVQLVQSGAEVKKPGESLKISCKGSGNGFTTYWIGWVRQVP GKGLEWMGIIYPDDSDTRYSPSFQGQVSISADKSISTAFLQWSSLKASDTAMYFCARLGDVET AMVGQDAFHIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG

[0412] VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKRGRKRRSGSGATNFSLLKQ AGDVEENPGPMVLQTQVFISLLLWISGAYGEIVLTQSPGTLSLSPGDRATLSCRASQSVSSSFL AWYQQKPGQAPRLLMYGASRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPF

[0413] TFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0414] SEQ ID NO:3 (NA) DMAb-FISW-033-HC

[0415] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0416] GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCAGCCTGAGGCTGT

[0417] CCTGCGCCGCCTCTACCTTCAACCTGTCTAATGCATGGATGAACTGGGTGCGGCAGGCAC

[0418] CAGGCAAGGGACTGGAGTGGGTGGGCCGCATCAAGAGCAAGACAGACGGCGGAACCAC

[0419] AGATTACGCCGCCCCTGTGAAGGGCAGGTTTATCATCAGCAGAGACGATTCCAAGAACA

[0420] TGCTGTATCTGCAGATGAATTCTCTGAAGATCGAGGACACCGCCGTGTACTATTGTGCCA

[0421] CAAATAAGGACAGCCTGGGCGGCTACGAGGCCTATGATATCTGGGGCCAGGGCACCATG

[0422] GTGACAGTGTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAG

[0423] TCTACCAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCA

[0424] GTGACCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTG

[0425] CTGCAGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTG

[0426] GGCACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAA

[0427] GAAGGTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTG AGCTGCTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGA

[0428] TCTCCCGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAG GTGAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAG

[0429] GGAGGAGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGG ACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCC

[0430] ATCGAGAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCT

[0431] GCCTCCAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGG

[0432] GCTTCTATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATT

[0433] ACAAGACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGA

[0434] CCGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGG

[0435] CCCTGCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0436] SEQ ID NO:4 (AA) DMAb-FISW-033-HC

[0437] MDWTWRILFLVAAATGTHAEVQLVESGGGLVKPGGSLRLSCAASTFNLSNAWMNWVRQAP

[0438] GKGLEWVGRIKSKTDGGTTDYAAPVKGRFIISRDDSKNMLYLQMNSLKIEDTAVYYCATNK

[0439] DSLGGYEAYDIWGQGTMVTVSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW

[0440] NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD

[0441] KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE

[0442] VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0443] SEQ ID NO: 5 (NA) DMAb-FISW-081-HC

[0444] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCCA

[0445] GGTGACACTGAGGGAGTCTGGACCAGCCCTGGTGAAGCCCACCCAGACACTGACCCTGA

[0446] CATGCACCTTCAGCGGCTTTTCTCTGAGCACCTCCGGCATGTGCGTGAGCTGGGTGAGGC

[0447] AGCCCCCTGGCAAGGCCCTGGAGTGGCTGGCCCTGATCGACTGGGACGATGGCAAGTAC

[0448] TATACCACAAGCCTGAAGGCCAGACTGACAATCTCTAAGGATACCAGCAAGAACCAGGT

[0449] GGTGCTGATCATGACAAATATGGACCCCGTGGATACAGCCACCTACTATTGCGCCCGCAC

[0450] ACTGTACGGCGACACCCTGTTCTATTTTGATTCCTGGGGCCAGGGCACCCTGGTGTCTGT

[0451] GAGCTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCTAC

[0452] CAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGTGA

[0453] CCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCTGC

[0454] AGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGGCA

[0455] CCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGAAG

[0456] GTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAGCTG

[0457] CTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATCTCC

[0458] CGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGTGAA

[0459] GTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGG

[0460] AGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGG CTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCATCGA

[0461] GAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTGCCTC

[0462] CAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGGCTTC

[0463] TATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTACAA

[0464] GACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGACCGT

[0465] GGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGCCCT

[0466] GCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAG

[0467] SEQ ID NO: 6 (AA) DMAb-FISW-081-HC

[0468] MDWTWRILFLVAAATGTHAQVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMCVSWVRQPP

[0469] GKALEWLALIDWDDGKYYTTSLKARLTISKDTSKNQVVLIMTNMDPVDTATYYCARTLYG

[0470] DTLFYFDSWGQGTLVSVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS

[0471] GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT

[0472] HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH

[0473] NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP

[0474] QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK

[0475] LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0476] SEQ ID NO:7 (NA) DMAb-FISW-085-HC

[0477] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0478] GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCAGCCTGAGGCTGT

[0479] CCTGCGCCGCCTCTGGCTTCAACAGCTCCAATGCATGGATGAAGTGGGTGCGGCAGGCA

[0480] CCAGGCAAGGGACTGGAGTGGGTGGGCCGCATCAAGAGCAAGACCGACTCCGGCACCA

[0481] CAGATTACGCCGCCCCTGTGAAGGGCAGGTTTACCATCTCTAGAGACGATAGCAAGAAC

[0482] ACACTGTATCTGCAGATGAATTCCCTGAAGTCTGAGGACACCGCCGTGTACTATTGTACC

[0483] ACAGACCTGGTGCGGTACTATGGCATGGACGTGTGGGGCCACGGCACCACAGTGACAGT

[0484] GTCTAGCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCTACC

[0485] AGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGTGAC

[0486] CGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCTGCA

[0487] GAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGGCAC

[0488] CCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGAAGG

[0489] TGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAGCTGC

[0490] TGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATCTCCC

[0491] GCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGTGAAG

[0492] TTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGA

[0493] GCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGC TGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCATCGAG

[0494] AAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTGCCTCC

[0495] AAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGGCTTCT

[0496] ATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTACAAG

[0497] ACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGACCGTG

[0498] GATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGCCCTG

[0499] CACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0500] SEQ ID NO:8 (AA) DMAb-FISW-085-HC

[0501] MDWTWRILFLVAAATGTHAEVQLVESGGGLVKPGGSLRLSCAASGFNSSNAWMKWVRQA

[0502] PGKGLEWVGRIKSKTDSGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLKSEDTAVYYCTTDL

[0503] VRYYGMDVWGHGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

[0504] SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK

[0505] THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV

[0506] HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE

[0507] PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS

[0508] KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0509] SEQ ID NO: 9 (NA) DMAb-FISW086-HC

[0510] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0511] GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGCAGCCAGGCAGGTCCCTGAGACTGT

[0512] CTTGCGCCGCCAGCGGCTTCACCTTTGACGATTACGCAATGCACTGGGTGAGGCAGGCAC

[0513] CTGGCAAGGGACTGGAGTGGGTGTCTGGCATCAGCTGGAACAGCGACTCCATCGTGTAC

[0514] GCCGATTCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAACGCCAAGAATTCTCTGTAT

[0515] CTGCAGATGAATAGCCTGCGGGCCGAGGATACAGCCTTCTACTATTGTGCCTCTATCGGC

[0516] TTTAGCTCCGGCTCTAGCGGACACTGGGGACAGGGCACCCTGGTGACAGTGTCCTCTGCC

[0517] AGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCTACCAGCGGCGGC

[0518] ACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGTGACCGTGAGCTGG

[0519] AACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCTGCAGAGCTCCGGC

[0520] CTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGGCACCCAGACATAT

[0521] ATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGAAGGTGGAGCCTAA

[0522] GAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAGCTGCTGGGCGGCCC

[0523] TAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATCTCCCGCACCCCTGA

[0524] GGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGTGAAGTTCAACTGGT

[0525] ACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAA

[0526] CTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCA AGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCATCGAGAAGACAATC

[0527] TCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTGCCTCCAAGCCGCGA

[0528] CGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGGCTTCTATCCATCCGA

[0529] TATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTACAAGACCACACCCC

[0530] CTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGACCGTGGATAAGTCTA

[0531] GGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGCCCTGCACAATCACT

[0532] ACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0533] SEQ ID NO: 10 (AA) DMAb-FISW086-HC

[0534] MDWTWRILFLVAAATGTHAEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAP

[0535] GKGLEWVSGISWNSDSIVYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAFYYCASIGFSS

[0536] GSSGHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL

[0537] TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC

[0538] PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK

[0539] TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT

[0540] LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD

[0541] KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0542] SEQ ID NO: 11 (NA) DMAb-FISW142-HC

[0543] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0544] GGTGCAGCTGGCCGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCTCCCTGAGGCTGT

[0545] CTTGCGCAGCAGCAGGCCTGAACAGCTCCAACGTGTGGATGAAGTGGGTGCGGCAGGCA

[0546] CCAGGCAAGGGACTGCAGTGGGTGGGCCGCATCAAGTCTAAGACCGACGGCGGAACCAC

[0547] AGATTATGCCGCCCCTGTGAGGGGCAGATTCACCATCAGCCGGGACGATTCCAAGAACA

[0548] CAGTGTACCTGCAGATGAATTCTCTGAAGGCCGAGGACACAGCCGTGTACTATTGTACCG

[0549] GCAGCAACACATACTATTACGGAATGGACGTGTGGGGACAGGGAACCACAGTGATCGTG

[0550] TCTAGCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCTACCA

[0551] GCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGTGACC

[0552] GTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCTGCAG

[0553] AGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGGCACC

[0554] CAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGAAGGT

[0555] GGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAGCTGCT

[0556] GGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATCTCCCG

[0557] CACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGTGAAGT

[0558] TCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAG

[0559] CAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGCT GAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCATCGAGA

[0560] AGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTGCCTCCA

[0561] AGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGGCTTCTA

[0562] TCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTACAAGA

[0563] CCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGACCGTGG

[0564] ATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGCCCTGC

[0565] ACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0566] SEQ ID NO: 12 (AA) DMAb-FISW142-HC

[0567] MDWTWRILFLVAAATGTHAEVQLAESGGGLVKPGGSLRLSCAAAGLNSSNVWMKWVRQA

[0568] PGKGLQWVGRIKSKTDGGTTDYAAPVRGRFTISRDDSKNTVYLQMNSLKAEDTAVYYCTGS

[0569] NTYYYGMDVWGQGTTVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW

[0570] NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD

[0571] KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE

[0572] VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0573] SEQ ID NO: 13 (NA) DMAb-FISW177-HC

[0574] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0575] GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCTCCCTGAGGCTGT

[0576] CTTGCGCCAGCTCCGGCTTCAACTTTAGCAATGCCTGGGTGAAGTGGGTGCGGCAGGCAC

[0577] CAGGCAAGGGACTGGAGTGGGTGGGACGCATCAAGTCCGAGACCGACGGCGGAACCAC

[0578] AGATTATGCCGCCCCTGTGAAGGGCCGCTTCACCATCAGCAGGGACAACTCCAGAAATA

[0579] CACTGTACCTGCAGCTGAACTCTCTGAGGACCGAGGATACAGGCATGTACTATTGTACCA

[0580] CATCTAGAAAGAGCTACTATTACCTGATGGACGTGTGGGGACAGGGAACCGCAGTGACA

[0581] GTGAGCGCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCT

[0582] ACCAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGT

[0583] GACCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCT

[0584] GCAGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGG

[0585] CACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGA

[0586] AGGTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAG

[0587] CTGCTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATC

[0588] TCCCGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGT

[0589] GAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGG

[0590] AGGAGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGAC TGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCAT

[0591] CGAGAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTG

[0592] CCTCCAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGG

[0593] CTTCTATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTA

[0594] CAAGACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGAC

[0595] CGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGC

[0596] CCTGCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0597] SEQ ID NO: 14 (AA) DMAb-FISW177-HC

[0598] MDWTWRILFLVAAATGTHAEVQLVESGGGLVKPGGSLRLSCASSGFNFSNAWVKWVRQAP

[0599] GKGLEWVGRIKSETDGGTTDYAAPVKGRFTISRDNSRNTLYLQLNSLRTEDTGMYYCTTSRK

[0600] SYYYLMDVWGQGTAVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN

[0601] SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK

[0602] THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV

[0603] HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE

[0604] PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS

[0605] KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0606] SEQ ID NO: 15 (NA) DMAb-FISW180-HC

[0607] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0608] GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCAGCCTGAGGCTGT

[0609] CCTGCGCAGCATCCGGCTTCTCTTTTACAAACGCATGGATGAAGTGGGTGCGGCAGGCAC

[0610] CAGGCAAGGGACTGGAGTGGGTGGGCCGCATCAAGTCCAAGACCGACGGCGGAACCAC

[0611] AGATTATACAGCCCCTGTGAAGGGCAGGTTCACCATCTCTAGAGACGATAGCAAGAACA

[0612] CAGTGTACCTGCAGATGAATTCTCTGAAGACCGAGGACACAGCCGTGTACTATTGTTCTG

[0613] GCAGCGGCCGGTACTATTACGGCATGGATATCTGGGGCCAGGGCACCACAGTGACCGTG

[0614] AGCTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCTACC

[0615] AGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGTGAC

[0616] CGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCTGCA

[0617] GAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGGCAC

[0618] CCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGAAGG

[0619] TGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAGCTGC

[0620] TGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATCTCCC

[0621] GCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGTGAAG

[0622] TTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGA

[0623] GCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGC TGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCATCGAG AAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTGCCTCC AAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGGCTTCT ATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTACAAG ACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGACCGTG GATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGCCCTG CACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0624] SEQ ID NO: 16 (AA) DMAb-FISW180-HC

[0625] MDWTWRILFLVAAATGTHAEVQLVESGGGLVKPGGSLRLSCAASGFSFTNAWMKWVRQAP GKGLEWVGRIKSKTDGGTTDYTAPVKGRFTISRDDSKNTVYLQMNSLKTEDTAVYYCSGSG RYYYGMDIWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0626] SEQ ID NO: 17 (NA) DMAb-FISW246-HC

[0627] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCTCTCTGAGGCTGA

[0628] GCTGCGCCGCCTCCGGCTTCAACCTGTCTAATGCATGGATGAACTGGGTGCGGCAGGCAC CAGGCAAGCGCCTGGAGTGGGTGGGCAGGATCAAGAGCAAGACCGACGGCGGAACCAC AGATTACGCCGCCCCTGTGAAGGGCAGGTTTACAATCTCTAGAGACGATAGCAAGAACA CCCTGTATCTGCAGACAAATTCCCTGAAGACCGAGGACACAGCCGTGTACTATTGCTGTA CCGACGCCGCCTACAACTATAATGATGGCGGCGGCGACTGGGATCACTGGGGACAGGGC ACCCTGGTGACAGTGAGCTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCT TCCTCTAAGTCTACCAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTC CCTGAGCCAGTGACCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTT

[0629] CCAGCCGTGCTGCAGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCC TCTAGCCTGGGCACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAG GTGGACAAGAAGGTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCC AGCACCTGAGCTGCTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACAC ACTGATGATCTCCCGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGA CCCCGAGGTGAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCA AGCCCAGGGAGGAGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTG CACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCC

[0630] AGCCCCCATCGAGAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTG

[0631] TACACCCTGCCTCCAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTG

[0632] GTGAAGGGCTTCTATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGA

[0633] GAACAATTACAAGACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTC

[0634] CAAGCTGACCGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGAT

[0635] GCACGAGGCCCTGCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGT GA

[0636] SEQ ID NO: 18 (AA) DMAb-FISW246-HC

[0637] MDWTWRILFLVAAATGTHAEVQLVESGGGLVKPGGSLRLSCAASGFNLSNAWMNWVRQA

[0638] PGKRLEWVGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQTNSLKTEDTAVYYCCTD

[0639] AAYNYNDGGGDWDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP

[0640] VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV

[0641] EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY

[0642] VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA

[0643] KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD

[0644] GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0645] SEQ ID NO: 19 (NA) DMAb-FISW298-HC

[0646] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCGA

[0647] GGTGCAGCTGGTGGAGAGCGGCGGCGGCCTGGTGAAGCCCGGCGGCAGCCTGAGGCTGT

[0648] CCTGCGCCGCCTCTGGCTTCTCTTTTAGCGACGCATGGATGACATGGGTGCGGCAGGGAC

[0649] CTGGAGAGGGACTGGAGTGGGTGGGCCGCATCAAGAGCAAGACCGACGGCGGAACCAC

[0650] AGATTACACAGCCGTGGTGAAGGGCAGGTTCACCATCAGCAGAGACGATTCCAAGCACA

[0651] CACTGTATCTGCAGATGAACTCCCTGAAGTCTGAGGATACCGCCGTGTACTATTGTACCA

[0652] CAGCCGGCTCCATGATGGAGGCCATCATCCCCTACGACGCCTTTGATATCTGGGGCCAGG

[0653] GCACCATGGTGACAGTGAGCTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCC

[0654] CTTCCTCTAAGTCTACCAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACT

[0655] TCCCTGAGCCAGTGACCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACAT

[0656] TTCCAGCCGTGCTGCAGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCT

[0657] CCTCTAGCCTGGGCACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCA

[0658] AGGTGGACAAGAAGGTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGT

[0659] CCAGCACCTGAGCTGCTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGAC

[0660] ACACTGATGATCTCCCGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAG

[0661] GACCCCGAGGTGAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGAC CAAGCCCAGGGAGGAGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGC

[0662] TGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTG

[0663] CCAGCCCCCATCGAGAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGG

[0664] TGTACACCCTGCCTCCAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTC

[0665] TGGTGAAGGGCTTCTATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCG

[0666] AGAACAATTACAAGACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATT

[0667] CCAAGCTGACCGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGA

[0668] TGCACGAGGCCCTGCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAG

[0669] TGA

[0670] SEQ ID NO: 20 (AA) DMAb-FISW298-HC

[0671] MDWTWRILFLVAAATGTHAEVQLVESGGGLVKPGGSLRLSCAASGFSFSDAWMTWVRQGP

[0672] GEGLEWVGRIKSKTDGGTTDYTAVVKGRFTISRDDSKHTLYLQMNSLKSEDTAVYYCTTAG

[0673] SMMEAIIPYDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT

[0674] VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP

[0675] KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD

[0676] GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG

[0677] QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF

[0678] FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0679] SEQ ID NO: 21 (NA) DMAb-LPAF021-HC

[0680] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCCA

[0681] GGTGACCCTGAGGGAGTCCGGACCAGCCCTGGTGAAGCCCACCCAGACACTGACCCTGA

[0682] CATGCACCTTCTCCGGCTTCTCTTTTACCACATCTGGAATGTGCGTGAGCTGGGTGCGGC

[0683] AGCCCCCTGGCAAGGCCCTGGAGTGGCTGGCCCTGATCGACTGGCGCGATAACAAGTTTT

[0684] ACTCTACAAGCCTGAGGACAAGACTGACCATCAGCAAGGACACCTCCAGGAATCAGGTG

[0685] GTGCTGACAATGACCGACATGGACCCCGTGGATACAGCCACCTATTCTTGCGCAAGGAT

[0686] GCTGTACGGCGACTTCGCAGGATACCTGGACCCCTGGGGACAGGGCACACTGGTGACCG

[0687] TGAGCTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCTAC

[0688] CAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGTGA

[0689] CCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCTGC

[0690] AGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGGCA

[0691] CCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGAAG

[0692] GTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAGCTG

[0693] CTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATCTCC

[0694] CGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGTGAA GTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGG

[0695] AGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGG

[0696] CTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCATCGA

[0697] GAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTGCCTC

[0698] CAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGGCTTC

[0699] TATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTACAA

[0700] GACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGACCGT

[0701] GGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGCCCT

[0702] GCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0703] SEQ ID NO: 22 (AA) DMAb-LPAF021-HC

[0704] MDWTWRILFLVAAATGTHAQVTLRESGPALVKPTQTLTLTCTFSGFSFTTSGMCVSWVRQPP

[0705] GKALEWLALIDWRDNKFYSTSLRTRLTISKDTSRNQVVLTMTDMDPVDTATYSCARMLYGD

[0706] FAGYLDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

[0707] ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH

[0708] TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN

[0709] AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ

[0710] VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL

[0711] TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0712] SEQ ID NO: 23 (NA) DMAb-LPAF-022-HC

[0713] ATGGACTGGACCTGGAGAATCCTGTTCCTGGTGGCCGCCGCCACCGGCACACACGCCCA

[0714] GGTGACACTGAGGGAGTCCGGACCAGCCCTGGTGAAGCCCACCCAGACACTGACCCTGA

[0715] CATGCACCTTCTCTGGCTTTTCTCTGAGCACATCCGGCATGTGCCTGACCTGGATCAGGA

[0716] GACCCCCTGGCAAGGCCCTGGAGTGGCTGGCCCTGATCGACTGGGACGATAACAAGTAC

[0717] TATAATACCTCCCTGCGGACACGCCTGACCATCTCTAAGGACACAAGCAAGAACCAGGT

[0718] GGTGCTGAAGGTGACCGACATGGACCCCGTGGATACAGGCACCTACTATTGCGCAAGGA

[0719] TGCTGTACGGCGACCTGGGCGGCCGGTTCGATCCATGGGGACAGGGCACACTGGTGACC

[0720] GTGAGCTCCGCCAGCACAAAGGGCCCTTCCGTGTTTCCCCTGGCCCCTTCCTCTAAGTCT

[0721] ACCAGCGGCGGCACAGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCTGAGCCAGT

[0722] GACCGTGAGCTGGAACTCCGGCGCCCTGACCTCTGGAGTGCACACATTTCCAGCCGTGCT

[0723] GCAGAGCTCCGGCCTGTACAGCCTGTCTAGCGTGGTGACAGTGCCCTCCTCTAGCCTGGG

[0724] CACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACCAAGGTGGACAAGA

[0725] AGGTGGAGCCTAAGAGCTGTGATAAGACCCACACATGCCCTCCCTGTCCAGCACCTGAG

[0726] CTGCTGGGCGGCCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAGGACACACTGATGATC

[0727] TCCCGCACCCCTGAGGTGACATGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAGGT GAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGG

[0728] AGGAGCAGTACAACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGAC

[0729] TGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGAGCAATAAGGCCCTGCCAGCCCCCAT

[0730] CGAGAAGACAATCTCCAAGGCAAAGGGACAGCCACGGGAGCCACAGGTGTACACCCTG

[0731] CCTCCAAGCCGCGACGAGCTGACCAAGAACCAGGTGTCCCTGACATGTCTGGTGAAGGG

[0732] CTTCTATCCATCCGATATCGCCGTGGAGTGGGAGTCTAATGGCCAGCCCGAGAACAATTA

[0733] CAAGACCACACCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTCTGTATTCCAAGCTGAC

[0734] CGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCCTGTTCTGTGATGCACGAGGC

[0735] CCTGCACAATCACTACACACAGAAGAGCCTGTCCCTGTCTCCAGGCAAGTGA

[0736] SEQ ID NO: 24 (AA) DMAb-LPAF-022-HC

[0737] MDWTWRILFLVAAATGTHAQVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMCLTWIRRPP

[0738] GKALEWLALIDWDDNKYYNTSLRTRLTISKDTSKNQVVLKVTDMDPVDTGTYYCARMLYG

[0739] DLGGRFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS

[0740] GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT

[0741] HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH

[0742] NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP

[0743] QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK

[0744] LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*

[0745] SEQ ID NO: 25 (NA) DMAb-FISW033-LC

[0746] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0747] GACATCCAGATGACCCAGTCTCCCAGCTCCCTGTCCGCCTCTGTGGGCGACAGGGTGACC

[0748] ATCACATGCCAGGCCAGCCAGGATATCACAAACTACCTGCACTGGTATCAGCAGAAGCC

[0749] CGGCAAGGCCCCTAAGCTGCTGATCTACGATGCAAGCTTCCTGGAGACCGGAGTGCCCA

[0750] GCAGGTTCAGCGGCTCCGGCTCTGGCACACACTTCACCTTTACAATCTCTAGCCTGCAGC

[0751] CAGAGGACATCGCCACCTACTATTGTCAGCAGTATGAGCACCTGCCATCCTTCGGCCCCG

[0752] GCACAAAGGTGGACTTCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCACCCA

[0753] GCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTCTACC

[0754] CTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTCTCAG

[0755] GAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCCTGAC

[0756] ACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACCAGG

[0757] GGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0758] SEQ ID NO: 26 (AA) DMAb-FISW033-LC MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQDITNYLHWYQQKPGK

[0759] APKLLIYDASFLETGVPSRFSGSGSGTHFTFTISSLQPEDIATYYCQQYEHLPSFGPGTKVDFKR

[0760] TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD

[0761] STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0762] SEQ ID NO: 27 (NA) DMAb-FISW085-LC

[0763] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0764] GACATCCAGATGACCCAGTCTCCTAGCTCCCTGTCCGCCTCTGTGGGCGACAGGGTGACC

[0765] ATCACATGCCAGGCCAGCCAGGATATCACCAACTACCTGAATTGGTATCAGCAGACACC

[0766] CGGCAAGGCCCCTAAGCTGCTGATCTACGACGCCAGCGATCTGAAGACCGGAGTGCCCT

[0767] CCAGGTTCAGCGGCTCCGGCTCTGGCACAGACTTTACCCTGACAATCTCTAGCCTGCAGC

[0768] CAGAGGACATCGCCACATATTTCTGTCAGCAGTTTGATTCCCTGCCCATCACCTTCGGCC

[0769] AGGGCACAAGACTGGATATCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCAC

[0770] CCAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTCT

[0771] ACCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTCT

[0772] CAGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCCT

[0773] GACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACC

[0774] AGGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0775] SEQ ID NO: 28 (AA) DMAb-FISW085-LC

[0776] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQDITNYLNWYQQTPGK

[0777] APKLLIYDASDLKTGVPSRFSGSGSGTDFTLTISSLQPEDIATYFCQQFDSLPITFGQGTRLDIKR

[0778] TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD

[0779] STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0780] SEQ ID NO: 29 (NA) DMAb-FISW086-LC

[0781] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0782] GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCTGTGGGCGACAGGGTGACC

[0783] ATCACATGCCAGGCCAGCCAGCACATCTCCGATTACCTGAACTGGTATCAGCAGAAGCC

[0784] CGGCAAGGCCCCTAAGCTGCTGATCTACGGCGCCTCTAATCTGGAGACCGGAGTGCCAA

[0785] GCAGGTTCAGCGGCTCCGGCTCTGGCACAGACTTCACCTTTACAATCTCTAGCCTGCAGC

[0786] CCGAGGACGTGGCCACATACTATTGTCAGCAGTATGATAACCTGCTGCTGACCTTTGGCC

[0787] AGGGCACAAGACTGGATATCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCAC

[0788] CCAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTCT

[0789] ACCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTCT

[0790] CAGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCCT GACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACC

[0791] AGGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0792] SEQ ID NO: 30 (AA) DMAb-FISW086-LC

[0793] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQHISDYLNWYQQKPGK

[0794] APKLLIYGASNLETGVPSRFSGSGSGTDFTFTISSLQPEDVATYYCQQYDNLLLTFGQGTRLDI

[0795] KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS

[0796] KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0797] SEQ ID NO: 31 (NA) DMAb-FISW142-LC

[0798] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0799] GACATCCAGATGACCCAGTCTCCTAGCTCCCTGTCCGCCTCTGTGGGCGACAGGGTGACC

[0800] ATCACATGCCAGGCCAGCCAGGATATCACAAACTACCTGAATTGGTATCAGCAGAAGCC

[0801] CGGCAAGGCCCCTAAGGTGCTGATCTACGACGCAAGCGATCTGCAGACCGGAGTGCCCT

[0802] CCAGATTCAGCGGCTCCGGCTCTGGCACAGATTTCACCTTTACAATCTCTAGCCTGCAGC

[0803] CAGAGGACATCGCCACCTACTATTGTCAGCAGTATGATGAGCTGCCACTGACCTTCGGCG

[0804] GCGGCACAAAGGTGGAGATCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCA

[0805] CCCAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTC

[0806] TACCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTC

[0807] TCAGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCC

[0808] TGACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCAC

[0809] CAGGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0810] SEQ ID NO: 32 (AA) DMAb-FISW142-LC

[0811] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQDITNYLNWYQQKPGK

[0812] APKVLIYDASDLQTGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDELPLTFGGGTKVEI

[0813] KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS

[0814] KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0815] SEQ ID NO: 33 (NA) DMAb-FISW177-LC

[0816] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0817] GACACCCAGCTGACACAGTCCCCCAGCTCCCTGTCTGCCAGCGTGGGCGACAGGGTGAC

[0818] CATCACATGCCAGGCCTCTCACGATATCACCAACTACCTGAATTGGTTCCAGCAGAAGCC

[0819] CGGCAAGAGCCCTAAGCTGCTGATCTATGATGCCTCCATCCTGGAGACCGGCGTGCCATC

[0820] TAGATTTTCCGGCTCTGGCAGCGGCACACACTTCACCTTTACAATCAACAATCTGCAGAG

[0821] CGAGGACATCGCCACATACTATTGTCAGCAGTTCGATTCCCTGCCACTGACCTTCGGCGG CGGCACAAAAGTGGGCCTGAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCACC

[0822] CAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTCTA

[0823] CCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTCTC

[0824] AGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCCTG ACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACCA GGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0825] SEQ ID NO: 34 (AA) DMAb-FlSW177-LC

[0826] MVLQTQVFISLLLWISGAYGDTQLTQSPSSLSASVGDRVTITCQASHDITNYLNWFQQKPGKS

[0827] PKLLIYDASILETGVPSRFSGSGSGTHFTFTINNLQSEDIATYYCQQFDSLPLTFGGGTKVGLKR TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0828] SEQ ID NO: 35 (NA) DMAb-FISW180-LC

[0829] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0830] GACATCCAGATGACCCAGTCTCCCAGCTCCCTGTCCGCCTCTGTGGGCGACAGGGTGACC

[0831] ATCACATGCCAGGCCAGCCAGGATATCACAAACTACCTGATCTGGTATCAGCAGAAGCC

[0832] CGGCAAGGCCCCTAAGCTGCTGATCTACGATGCCAGCAATCTGGAGACCGGCGTGCCTTC

[0833] CAGATTCAGCGGCTCCGGCTCTGGCACAGACTTCACCTTTACAATCTCTAGCCTGCAGCC

[0834] AGAGGATATCGCCACCTACTATTGTCAGCAGTTCGAGGCCCTGCCAATCACCTTTGGCCC

[0835] CGGCACAAAGGTGGACATCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCACC

[0836] CAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTCTA

[0837] CCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTCTC

[0838] AGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCCTG ACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACCA GGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0839] SEQ ID NO: 36 (AA) DMAb-FISW180-LC

[0840] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQDITNYLIWYQQKPGKA

[0841] PKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQFEALPITFGPGTKVDIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDS TYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0842] SEQ ID NO: 37 (NA) DMAb-FISW246-LC

[0843] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0844] GACATCCAGATGACCCAGTCTCCTAGCTCCCTGTCTGCCAGCGTGGGCGACAGGGTGACC ATCACATGCCAGGCCAGCCAGGATATCACAAACTACCTGAATTGGTATCAGCAGAAGCC

[0845] CGGCAAGGCCCCTAAGCTGCTGATCTACGATGCCTCCAACCTGGAGACCGGCGTGCCCTC

[0846] TAGATTCTCCGGCTCTGGCAGCGTGACCGACTTCGCCTTTACAATCAGCGGCCTGCAGCC

[0847] AGAGGATATCGGCACATACTATTGTCAGCAGTATGAGTCCCTGCCACTGACCTTCGGCGG

[0848] CGGCACAAAGGTGGAGATCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCACC

[0849] CAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTCTA

[0850] CCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTCTC

[0851] AGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCCTG

[0852] ACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACCA

[0853] GGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0854] SEQ ID NO: 38 (AA) DMAb-FISW246-LC

[0855] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQDITNYLNWYQQKPGK

[0856] APKLLIYDASNLETGVPSRFSGSGSVTDFAFTISGLQPEDIGTYYCQQYESLPLTFGGGTKVEIK

[0857] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK

[0858] DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0859] SEQ ID NO: 39 (NA) DMAb-FISW298-LC

[0860] ATGGTGCTGCAGACCCAGGTGTTCATCAGCCTGCTGCTGTGGATCTCCGGCGCCTACGGC

[0861] GACATCCAGATGACCCAGTCTCCTAGCTCCCTGTCCGCCTCTGTGGGCGACAGGGTGACC

[0862] ATCACATGCCAGGCCAGCCAGGATATCACAAACTACCTGCACTGGTATCAGCAGAAGCC

[0863] CGGCAAGGCCCCTAAGCTGCTGATCTACGACGCCAGCAATCTGGAGACCGGAGTGCCCT

[0864] CCAGATTCAGCGGCTCCGGCTCTGGCACAGATTTCACCTTTACAATCTCTAGCCTGCAGC

[0865] CAGAGGACCTGGCCACCTACTATTGTCAGCAGTATGATAACCTGCCCCTGACCTTCGGCG

[0866] GCGGCACAAAGGTGGAGATCAAGCGTACAGTGGCCGCCCCCAGCGTGTTCATCTTTCCA

[0867] CCCAGCGACGAGCAGCTGAAGTCCGGCACCGCCTCTGTGGTGTGCCTGCTGAACAATTTC

[0868] TACCCTCGGGAGGCCAAGGTGCAGTGGAAGGTGGATAACGCCCTGCAGTCCGGCAATTC

[0869] TCAGGAGAGCGTGACCGAGCAGGACTCCAAGGATTCTACATATAGCCTGAGCTCCACCC

[0870] TGACACTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCAC

[0871] CAGGGGCTGTCAAGTCCAGTCACTAAAAGTTTCAATAGGGGAGAATGTTGA

[0872] SEQ ID NO: 40 (AA) DMAb-FlSW298-LC

[0873] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGDRVTITCQASQDITNYLHWYQQKPGK

[0874] APKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDLATYYCQQYDNLPLTFGGGTKVEI

[0875] KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS

[0876] KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* SEQ ID NO: 41 (NA) DMAb-FISW081-LC

[0877] ATGGCCTGGACCCCTCTGCTGCTGCCTCTGCTGACCTTCTGCACAGTGAGCGAGGCCTCC

[0878] AGCTACGTGCTGACACAGCCCCCTAGCATGTCCGTGGCACCAGGCAAGACAGCAAGGAT

[0879] CACCTGCGGCGGAAACAATATCGGCTCCAAGAACGTGCACTGGTATCAGCAGAAGCCAG

[0880] GACAGGCACCAGTGGTGGTCATCTACTATGACTCTGCCAGGCCTAGCGGCATCCCAGAG

[0881] AGATTCTCTGGCAGCAACTCCGGCAATACCGCCACACTGACCATCTCCCGCGTGGAGGCA

[0882] GGCGACGAGGCAGATTACTATTGTCAAGTGTGGGACTCTACCACAGATCACTACGTGTTT

[0883] GGCATCGGCACAAAGGTGACCGTGCTGGGATCCGCCGCCACCATGGCCTGGACCCCTCT

[0884] GCTGCTGCCTCTGCTGACCTTCTGCACAGTGAGCGAGGCCTCCGGACAGCCAAAGGCAG

[0885] CACCATCTGTGACCCTGTTCCCTCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACC

[0886] CTGGTGTGCCTGATCTCCGACTTTTACCCAGGAGCAGTGACAGTGGCATGGAAGGCCGAT

[0887] TCTAGCCCTGTGAAGGCCGGCGTGGAGACCACAACCCCATCTAAGCAGAGCAACAATAA

[0888] GTACGCCGCCTCCTCTTATCTGTCCCTGACCCCCGAGCAGTGGAAGTCTCACAGGAGCTA

[0889] TTCCTGCCAGGTGACACACGAGGGCAGCACAGTGGAGAAGACCGTGGCCCCTACAGAGT GTTCCTGA

[0890] SEQ ID NO: 42 (AA) DMAb-FISW081-LC

[0891] MAWTPLLLPLLTFCTVSEASSYVLTQPPSMSVAPGKTARITCGGNNIGSKNVHWYQQKPGQA

[0892] PVVVIYYDSARPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSTTDHYVFGIGTKV

[0893] TVLGSAATMAWTPLLLPLLTFCTVSEASGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYP

[0894] GAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTV

[0895] EKTVAPTECS*

[0896] SEQ ID NO: 43 (NA) DMAb-LPAF021-LC

[0897] ATGGCCTGGACCCCTCTGCTGCTGCCTCTGCTGACCTTCTGCACAGTGAGCGAGGCCTCC

[0898] TCCTACGAGCTGACACAGGCCCTGAGCGTGTCCGTGGCCCTGGGACAGACAGCAAGGAT

[0899] CACCTGCGGCGGAAACAATATCGGCTACAAGAACGTGCACTGGTACCAGTATAAGCCAG

[0900] GACAGGCACCCGTGCTGGTCATCTACAGGGACACCAATAGACCATCTGGCATCCCCGAG

[0901] AGGTTCAGCGGCAGCAACTCCGGCAATACCGCCACACTGACCATCAGCGGAGCACAGGG

[0902] CGGCGACGAGGCAGATTACTATTGTCAAGTGTGGGATAGCTCCTCTGTGGCCTTCGGCGG

[0903] CGGCACAAAGCTGACCGTGCTGGGATCCGCCGCCACCATGGCCTGGACCCCTCTGCTGCT

[0904] GCCTCTGCTGACCTTCTGCACAGTGAGCGAGGCCTCCGGACAGCCAAAGGCAGCACCAT

[0905] CTGTGACCCTGTTCCCTCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGT

[0906] GCCTGATCTCCGACTTTTACCCAGGAGCAGTGACAGTGGCATGGAAGGCCGATTCTAGCC

[0907] CTGTGAAGGCCGGCGTGGAGACCACAACCCCATCTAAGCAGAGCAACAATAAGTACGCC GCCTCCTCTTATCTGTCCCTGACCCCCGAGCAGTGGAAGTCTCACAGGAGCTATTCCTGC

[0908] CAGGTGACACACGAGGGCAGCACAGTGGAGAAGACCGTGGCCCCTACAGAGTGTTCCTG A

[0909] SEQ ID NO: 44 (AA) DMAb-LPAF021-LC

[0910] MAWTPLLLPLLTFCTVSEASSYELTQALSVSVALGQTARITCGGNNIGYKNVHWYQYKPGQ

[0911] APVLVIYRDTNRPSGIPERFSGSNSGNTATLTISGAQGGDEADYYCQVWDSSSVAFGGGTKLT

[0912] VLGSAATMAWTPLLLPLLTFCTVSEASGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPG

[0913] AVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVE KTVAPTECS*

[0914] SEQ ID NO: 45 (NA) DMAb-LPAF022-LC

[0915] ATGGCCTGGACCCCTCTGCTGCTGCCTCTGCTGACCTTCTGCACAGTGAGCGAGGCCTCC

[0916] TCCTACGAGCTGACCCAGCCCCTGAGCGTGTCCGTGGCCCTGGGACAGACCGCAAGGAT

[0917] CACATGCGGCGGAAACAATATCGGCATCAAGAACGTGCACTGGTACCAGCAGAGGCCAG

[0918] GACAGGCACCCGTGCTGGTCATCTACACAGACATCAATAGGCCATCTGGCATCCCCGAG

[0919] AGATTCTCTGGCAGCAACTCCGGCAATACCGCCACACTGATCATCTCTAAGGCACAGGCA

[0920] GGCGACGAGGCAGATTATTTCTGTCAAGTGTGGGATAGCAACACCGTGGTGTTTGGCGG

[0921] CGGAACCAAGCTGACAGTGCTGGGATCCGCCGCCACCATGGCCTGGACCCCTCTGCTGCT

[0922] GCCTCTGCTGACCTTCTGCACAGTGAGCGAGGCCTCCGGACAGCCAAAGGCAGCACCAT

[0923] CTGTGACCCTGTTCCCTCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGT

[0924] GCCTGATCTCCGACTTTTACCCAGGAGCAGTGACAGTGGCATGGAAGGCCGATTCTAGCC

[0925] CTGTGAAGGCCGGCGTGGAGACCACAACCCCATCTAAGCAGAGCAACAATAAGTACGCC

[0926] GCCTCCTCTTATCTGTCCCTGACCCCCGAGCAGTGGAAGTCTCACAGGAGCTATTCCTGC

[0927] CAGGTGACACACGAGGGCAGCACAGTGGAGAAGACCGTGGCCCCTACAGAGTGTTCCTG A

[0928] SEQ ID NO: 46 (AA) DMAb-LPAF022-LC

[0929] MAWTPLLLPLLTFCTVSEASSYELTQPLSVSVALGQTARITCGGNNIGI

[0930] KNVHWYQQRPGQAPVLVIYTDINRPSGIPERFSGSNSGNTATLIISKAQAGDEADYFC

[0931] QVWDSNTVVFGGGTKLTVLGSAATMAWTPLLLPLLTFCTVSEASGQPKAAPSVTLF

[0932] PPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAAS SYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS*

[0933] SEQ ID NO:47 - Amino acid sequence of AF9C-HC MDWTWRILFLVAAATGTHAEVQLVESGAELKKPGSSVRVSCKASRHT FSSYSITWVRQAPGQGLEWMGE11PIFGTANYGQKFQGRVT1TADASTSTAY1EVTSLR FEDTAIYYCARDLAPYGDRFYFHYGMDVWGQGTLVTVSSASTKGPSVFPLAPSSKST SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGK*

[0934] SEQ ID NO:48 - Nucleic acid sequence of AF9C-HC atggactggacctggagaatcctgttcctggtggccgccgccaccggcacacacgccgaggtgcagctggtg gagtccggagcagagctgaagaagccaggcagctccgtgagggtgtcttgcaaggccagcagacacacctctctagctactctatc acatgggtgaggcaggcaccaggacagggactggagtggatgggcgagatcatccctatctcggcaccgccaactacggccaga agttcagggcagggtgaccatcacagccgacgccagcacctccacagcctatatcgaggtgaccagcctgcggttgaggatacag ccatctactattgtgcccgggacctggcaccatatggcgatcgcttctactttcactatggaatggacgtgtggggacagggcaccctg gtgacagtgtcctctgccagcacaaagggcccttccgtgtttcccctggcccctcctctaagtctaccagcggcggcacagccgccct gggatgtctggtgaaggattacttccctgagccagtgaccgtgagctggaactccggcgccctgacctctggagtgcacacattcca gccgtgctgcagagctccggcctgtacagcctgtctagcgtggtgacagtgccctcctctagcctgggcacccagacatatatctgca acgtgaatcacaagccctctaataccaaggtggacaagaaggtggagcctaagagctgtgataagacccacacatgccctccctgtc cagcacctgagctgctgggcggccctagcgtgttcctgtttccacccaagccaaaggacacactgatgatctcccgcacccctgaggt gacatgcgtggtggtggacgtgtctcacgaggaccccgaggtgaagttcaactggtacgtggatggcgtggaggtgcacaatgcca agaccaagcccagggaggagcagtacaactctacctatagagtggtgagcgtgctgacagtgctgcaccaggactggctgaacggc aaggagtataagigcaaggtgagcaataaggccctgccagcccccatcgagaagacaatctccaaggcaaagggacagccacgg gagccacaggtgtacaccctgcctccaagccgcgacgagctgaccaagaaccaggtgtccctgacatgtctggtgaagggctctat ccatccgatatcgccgtggagtgggagtctaatggccagcccgagaacaattacaagaccacaccccctgtgctggactctgatggc agcttcttctgtattccaagctgaccgtggataagtctaggiggcagcagggcaacgtgtttcctgttctgtgatgcacgaggccctgc acaatcactacacacagaagagcctgtccctgtctccaggcaagtga

[0935] SEQ ID NO:49 Amino acid sequence of AF9C-LC

[0936] MVLQTQVFISLLLWISGAYGDIQLTQSPSSLSASVGDGVTITCRASQDIS NSLAWYQQKPGKAPNLLISAASTLQSGVPSRFSGSGSGTIFTLTISSLQPEDFATYYCQ QLNNYPFTFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC*

[0937] SEQ ID NO:50 Nucleic acid sequence of AF9C-LC atggtgctgcagacccaggtgtcatcagcctgctgctgtggatctccggcgcctacggcgacatccagctgac acagtcccctagctccctgtccgcctctgtgggcgacggagtgaccatcacatgcagggccagccaggatatcagcaactccctggc ctggtaccagcagaagcccggcaaggcccctaatctgctgatctctgccgcaagcaccctgcagtccggagtgccctctagatctct ggcagcggctccggcacaatctttaccctgacaatctctagcctgcagccagaggatttcgccacctactatgtcagcagctgaacaa ttatcccttcacctttggccagggcacaaggctggagatcaagcgtacagtggccgcccccagcgtgttcatctttccacccagcgacg agcagctgaagtccggcaccgcctctgtggtgtgcctgctgaacaattctaccctcgggaggccaaggtgcagtggaaggtggata acgccctgcagtccggcaattctcaggagagcgtgaccgagcaggactccaaggatctacatatagcctgagctccaccctgacact gagcaaggccgactacgagaagcacaaggtgtatgcctgtgaggtcacccaccaggggctgtcaagtccagtcactaaaagtttcaa taggggagaatgttga

[0938] SEQ ID NO:51 Amino acid sequence of AG7C-HC

[0939] MDWTWRILFLVAAATGTHAQVQLQESGPGLVKPSQTLSLTCAVSGDS ISDGPYYWSWIRQYPGKGLEWIGYFYYSSTTYFNPSLKSRISMSVDTSKNHFSLKVNS VTAADTAVYYCARDLEGHTFHDWGHGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGK*

[0940] SEQ ID NO:52 Nucleic acid sequence of AG7C-HC atggactggacctggagaatcctgttcctggtggccgccgccaccggcacacacgcccaggtgcagctgcag gagtctggaccaggactggtgaagcctcccagaccctgtctctgacatgcgccgtgagcggcgactccatctctgatggcccatact atggtcctggatcaggcagtaccccggcaagggcctggagtggatcggctacttctactatagctccaccacatactcaacccctctc tgaagagcagaatcagcatgtccgtggacaccagcaagaaccacttctccctgaaggtcaattctgtgaccgccgccgatacagccgt gtactattgtgccagggacctggagggccacacatttcacgattggggccacggcaccctggtgacagtgtctagcgccagcacaaa gggccctccgtgttcccctggcccctcctctaagtctaccagcggcggcacagccgccctgggatgtctggtgaaggatactcc ctgagccagtgaccgtgagctggaactccggcgccctgacctctggagtgcacacattccagccgtgctgcagagctccggcctgt acagcctgtctagcgtggtgacagtgccctcctctagcctgggcacccagacatatatctgcaacgtgaatcacaagccctctaatacc aaggtggacaagaaggtggagcctaagagctgtgataagacccacacatgccctccctgtccagcacctgagctgctgggcggccc tagcgtgttcctgttccacccaagccaaaggacacactgatgatctcccgcacccctgaggtgacatgcgtggtggtggacgtgtctc acgaggaccccgaggtgaagttcaactggtacgtggatggcgtggaggtgcacaatgccaagaccaagcccagggaggagcagt acaactctacctatagagtggtgagcgtgctgacagtgctgcaccaggactggctgaacggcaaggagtataagtgcaaggtgagca ataaggccctgccagcccccatcgagaagacaatctccaaggcaaagggacagccacgggagccacaggtgtacaccctgcctcc aagccgcgacgagctgaccaagaaccaggtgtccctgacatgtctggtgaagggctctatccatccgatatcgccgtggagtggga gtctaatggccagcccgagaacaattacaagaccacaccccctgtgctggactctgatggcagctcttctgtattccaagctgaccgt ggataagtctaggtggcagcagggcaacgtgttttcctgtctgtgatgcacgaggccctgcacaatcactacacacagaagagcctgt ccctgtctccaggcaagtga

[0941] SEQ ID NO:53 Amino acid sequence of AG7C-LC

[0942] MVLQTQVFISLLLWISGAYGDIQMTQSPSSLSASVGERVTITCRASQIIG NYLNWYQQRPGEAPKLLIYTTSNLQSGVPSRFSGAVSQTDFTLTIRGLQREDFATYY CQQSHSAPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC*

[0943] SEQ ID NO:54 Nucleic acid sequence of AG7C-LC

[0944] Atggtgctgcagacccaggtgtcatcagcctgctgctgtggatctccggcgcctacggcgacatccagatga cccagtcccccagctccctgtctgccagcgtgggagagagggtgaccatcacatgcagggcctctcagatcatcggcaactacctga attggtatcagcagaggccaggagaggcacctaagctgctgatctacaccacatctaacctgcagagcggagtgccaagccggtct ccggagccgtgagccagaccgactttaccctgacaatcaggggcctgcagagagaggatttcgccacatactattgtcagcagtccc actctgccccctatacctttggccagggcacaaaggtggagatcaagcgtacagtggccgcccccagcgtgtcatcttccacccag cgacgagcagctgaagtccggcaccgcctctgtggtgtgcctgctgaacaatttctaccctcgggaggccaaggtgcagtggaaggt ggataacgccctgcagtccggcaatctcaggagagcgtgaccgagcaggactccaaggattctacatatagcctgagctccaccct gacactgagcaaggccgactacgagaagcacaaggtgtatgcctgtgaggtcacccaccaggggctgtcaagtccagtcactaaaa gttcaataggggagaatgtga

[0945] SEQ ID NO: 55 nucleic acid sequence of pVACC-NPH1atggactggacctggattctgttcctggtggccgccgccacaagagtgcactctatggccagccagggcaccaagcggtcctacgag cagatggagacaggcggcgagcgccaggataccacagagatcagggcatctgtgggaaggatgatcggcggaatcggcaggttct acatccagatgtgcaccgagctgaagctgtctgactatgatggccgcctgatccagaactctatcacaatcgagaggatggtgctgag cgcctttgacgagaggagaaataagtatctggaggagcaccctagcgccggcaaggaccccaagaagaccggcggcccaatctac aggaggatcgacggcaagtggacaagggagctgatcctgtatgataaggaggagatcaggagagtgtggagacaggcaaacaat ggagaggacgcaaccgcaggactgacacacatcatgatctggcactccaacctgaatgacgcaacctaccagaggacacgcgccct ggtgagaaccggaatggacccccggatgtgcagcctgatgcagggctccacactgcctcggcgcagcggagcagcaggagcagcc gtgaagggcgtgggcaccatcgccatggagctgatccggatgatcaagcgcggcatcaacgacaggaatttctggagaggcgagaa cggcaggagaaccagggtggcctatgagagaatgtgcaatatcctgaagggcaagtttcagacagccgcccagagagccatgatgg accaggtgagggagtctagaaaccccggcaatgccgagatcgaggatctgatcttcctggcacgcagcgccctgatcctgaggggct ctgtggcacacaagagctgcctgccagcctgcgtgtacggactggcagtggcaagcggacacgacttcgagcgggagggctattcc ctggtgggcatcgatccctttaagctgctgcagaactcccaggtggtgtctctgatgcgcccaaacgagaatcccgcccacaagtccc agctggtgtggatggcctgtcactctgccgccttcgaggacctgagagtgagctcctttatccggggcaagaaagtgatccctcgcgg caagctgtccaccaggggcgtgcagatcgcctctaacgagaatgtggagaccatggatagcaacacactggagctgaggtccagat actgggcaatcaggacccgcagcggcggcaacacaaatcagcagaaggcatccgccggacagatctctgtgcagcctaccttctcc gtgcagagaaatctgccatttgagagggcaacagtgatggcagccttcagcggcaacaatgagggccggacctccgacatgcgcac agaagtgatcaggatgatggagtctgccaagccagaggatctgagcttccagggccggggcgtgtttgagctgtccgacgagaagg ccaccaaccccatcgtgcctagctttgatatgtccaatgagggctcttacttctttggcgacaacgccgaggagtatgataattga

[0946] SEQ ID NO: 56 amino acid sequence of pVACC-NPH1

[0947] MDWTWILFLVAAATRVHSMASQGTKRSYEQMETGGERQDTTEIRAS VGRMIGGIGRFYIQMCTELKLSDYDGRLIQNSITIERMVLSAFDERRNKYLEEHPSAG KDPKKTGGPIYRRIDGKWTRELILYDKEEIRRVWRQANNGEDATAGLTHIMIWHSNL NDATYQRTRALVRTGMDPRMCSLMQGSTLPRRSGAAGAAVKGVGTIAMELIRMIK RGINDRNFWRGENGRRTRVAYERMCNILKGKFQTAAQRAMMDQVRESRNPGNAEI EDLIFLARSALILRGSVAHKSCLPACVYGLAVASGHDFEREGYSLVGIDPFKLLQNSQ VVSLMRPNENPAHKSQLVWMACHSAAFEDLRVSSFIRGKKVIPRGKLSTRGVQIASN ENVETMDSNTLELRSRYWAIRTRSGGNTNQQKASAGQ1SVQPTFSVQRNLPFERATV MAAFSGNNEGRTSDMRTEVIRMMESAKPEDLSFQGRGVFELSDEKATNPIVPSFDMS NEGSYFFGDNAEEYDN*

[0948] SEQ ID NO: 57 nucleic acid sequence of pVACC-NPH3atggactggacctggattctgtcctggtggcagcagcaacacgcgtgcactccatggcctctcagggcaccaa gaggagctacgagcagatggagaccgacggcgatagacagaacgccacagagatccgggcctccgtgggcaagatgatcgacg gcatcggccgcttctatatccagatgtgcacagagctgaagctgtccgatcacgagggccggctgatccagaactccctgaccatcga gaagatggtgctgtctgcctttgacgagaggagaaataagtacctggaggagcaccctctgccggcaaggaccccaagaagaccg gcggcccaatctacaggagggtggacggcaagtggatgagagagctggtgctgtatgataaggaggagatcaggagaatctggcg gcaggccaacaatggagaggacgccacctccggactgacacacatcatgatctggcactctaacctgaatgacgccacctatcagag gacaagagccctggtgagaacaggcatggacccccggatgtgctctctgatgcagggcagcaccctgccacggcgctccggagca gcaggagcagccgtgaagggcatcggcaccatggtcatggagctgatccggatggtgaagcgcggcatcaacgataggaattctg gagaggcgagaacggaaggaagacacgcagcgcctacgagcgcatgtgcaatatcctgaagggcaagttcagaccgcagcaca gagggcaatggtggaccaggtgagggagtccagaaaccccggcaatgccgagatcgaggatctgatcttcctggcacgctctgccc tgatcctgaggggctccgtggcacacaagtctgcctgcctgcatgtgcatacggaccagccgtgagctccggatacgactcgagaa ggagggctatccctggtgggcatcgatccttttaagctgctgcagaacagccagatctatccctgatcagaccaaacgagaatcccg cccacaagagccagctggtgtggatggcctgtcactccgccgccttcgaggacctgagactgctgtctttatccggggcacaaaggt gtctccacgcggcaagctgagcaccaggggcgtgcagatcgccagcaacgagaatatggataatatgggctctagcacactggagc tgaggtctggctatgggcaatcaggacccgcagcggcggcaacacaaatcagcagagagcaagcgccggacagacctccgtgca gcccacattcagcgtgcagcggaacctgcctttgagaagtccacaatcatggccgcctcaccggcaatacagagggacggaccag cgacatgagggcagagatcatcaggatgatggagggcgccaagcctgaggaggtgtctttcaggggcagaggcgtgttgagctga gcgacgagaaggccaccaaccccatcgtgcctcttgatatgagcaatgagggctcctactcttggcgacaacgccgaggagtat gataattga

[0949] SEQ ID NO: 58 amino acid sequence of pVACC-NPH3

[0950] MDWTWILFLVAAATRVHSMASQGTKRSYEQMETDGDRQNATEIRAS

[0951] VGKMIDGIGRFYIQMCTELKLSDHEGRLIQNSLTIEKMVLSAFDERRNKYLEEHPSAG KDPKKTGGPIYRRVDGKWMRELVLYDKEEIRRIWRQANNGEDATSGLTHIMIWHSN LND ATYQRTRALVRTGMDPRMC SLMQGSTLPRRS GAAGAAVKGIGTMVMELIRMV KRGINDRNFWRGENGRKTRSAYERMCNILKGKFQTAAQRAMVDQVRESRNPGNAE 1EDLIFLARSAL1LRGSVAHKSCLPACAYGPAVSSGYDFEKEGYSLVG1DPFKLLQNSQ IYSLIRPNENPAHKSQLVWMACHSAAFEDLRLLSFIRGTKVSPRGKLSTRGVQIASNE NMDNMGSSTLELRSGYWAIRTRSGGNTNQQRASAGQTSVQPTFSVQRNLPFEKSTI MAAFTGNTEGRTSDMRAEIIRMMEGAKPEEVSFRGRGVFELSDEKATNPIVPSFDMS NEGSYFFGDNAEEYDN*

[0952] SEQ ID NO: 59 nucleic acid sequence of CaO9 HA (pHAH1) atggactggacctggattctgttcctggtggcagcagcaacacgggtgcacagcatgaaggccatcctggtggt gctgctgtacacctcgccacagccaacgccgataccctgtgcatcggctatcacgccaacaatctaccgacacagtggatacagtg ctggagaagaatgtgaccgtgacacacagcgtgaacctgctggaggacaagcacaatggcaagctgtgcaagctgcgcggcgtgg cccctctgcacctgggcaagtgcaacatcgccggctggattctgggcaatcctgagtgtgagtctctgagcaccgccagctcctggag ctacatcgtggagaccccctctagcgataacggcacatgctaccctggcgactttatcgatatgaggagctgcgggagcagctgagc agcgtgagcagctcgagaggtcgagatcttccccaagacctctagctggcctaaccacgactccaataagggagtgacagcagca tgtcctcacgcaggcgccaagagctctacaagaacctgatctggctggtgaagaagggcaatcctacccaaagctgtccaagtcta tatcaacgataagggcaaggaggtgctggtgctgtggggcatccaccacccaagcacctccgccgaccagcagtccctgtaccaga atgccgatgcctacgtgttcgtgggctcctctaggtactctaagaagttcaagccagagatcgccatcaggcccaaggtgagagacca ggagggcagaatgaactactattggaccctggtggagcctggcgataagatcacctttgaggccacaggcaacctggtggtgccaag atatgccttcgccatggagagaaatgccggcagcggcatcatcatctccgacaccccagtgcacgattgcaacaccacatgtcagac ccccaagggcgccatcaacacaagcctgcctttccagaatatccacccaatcacaatcggcaagtgcccaaagtacgtgaagtccac caagctgaggctggcaacaggactgcgcaatatcccctctatccagagcaggggcctgtttggagcaatcgcaggcttcatcgaggg cggctggaccggaatggtggacggctggtacggctatcaccaccagaacgagcagggctccggatatgcagcagacctgaagtct acccagaatgccatcgatgagatcacaaacaaggtcaattccgtgatcgagaagatgaacacccagtttacagccgtgggcaaggag ttcaatcacctggagaagcggatcgagaacctgaataagaaggtggacgatggctttctggatatctggacctacaacgccgagctgc tggtgctgctggagaatgagcggacactggactaccacgatagcaacgtgaagaatctgtatgagaaggtgcgctcccagctgaaga acaatgccaaggagatcggcaacggctgcttcgagttttaccacaagtgcgacaacacctgtatggagtccgtgaagaatggcacata cgattatcctaagtattctgaggaggccaagctgaatagggaggagatcgacggcgtgaagctggagtctaccagaatctaccagatc ctggccatctatagcacagtggccagctccctggtgctggtggtgtctctgggcgccatcagcttctggatgtgctccaacggctctctg cagtgcaggatctgtatctgatgataa

[0953] SEQ ID NO: 60 amino acid sequence of Ca09 HA (pHAH1) MDWTWILFLVAAATRVHSMKAILVVLLYTFATANADTLCIGYHANNSTDTVDTVLEK NVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYI VETPSSDNGTCYPGDFIDYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPH AGAKSFYKNLIWLVKKGNSYPKLSKSYINDKGKEVLVLWGIHHPSTSADQQSLYQNA DAYVFVGSSRYSKKFKPEIAIRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRY AFAMERNAGSGIIISDTPVHDCNTTCQTPKGAINTSLPFQNIHPITIGKCPKYVKSTKLR LATGLRNIPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQ NAIDEITNKVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELL VLLENERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCMESVKNG TYDYPKYSEEAKLNREEIDGVKLESTRIYQILAIYSTVASSLVLVVSLGAISFWMCSNG SLQCRICI*

[0954] SEQ ID NO: 61 (dominant peptide 86)

[0955] AHKSQLVWMAC

[0956] SEQ ID NO: 62 (dominant peptide 91) VSSFIRGKKVI

[0957] SEQ ID NO: 63 (dominant peptide 95)

[0958] ASNENVETMDSN

[0959] SEQ ID NO: 64 (dominant peptide 102)

[0960] AIRTRS GGNTNQQKA

[0961] SEQ ID NO: 65 (dominant peptide 107)

[0962] TFSVQRNLPFE

[0963] SEQ ID NO: 66 (dominant peptide 109)

[0964] PFERATVMAAF

[0965] SEQ ID NO: 67 (dominant peptide 115)

[0966] DMRTEVIRMME

[0967] SEQ ID NO: 68 (dominant peptide 120)

[0968] QGRGVFELSDE

[0969] SEQ ID NO: 69 (dominant peptide 53)

[0970] MELIRMVKRGINDRN

[0971] SEQ ID NO: 70 (dominant peptide 65) VDQVRESRNPGNAEI

[0972] SEQ ID NO: 71 (dominant peptide 89)

[0973] SAAFEDLRLLSFIRG

[0974] SEQ ID NO: 72 (dominant peptide 113)

[0975] GNTEGRTSDMRAEII

[0976] SEQ ID NO: 73 (dominant peptide 59)

[0977] TRSAYERMCNILKGK

[0978] SEQ ID NO: 74 (dominant peptide 71)

[0979] RGSVAHKSCLPACAY

[0980] SEQ ID NO: 75 (dominant peptide 95)

[0981] STRGVQIASNENMDN

[0982] SEQ ID NO: 76 (dominant peptide 119)

[0983] EVSFRGRGVFELSDE

[0984] SEQ ID NO: 77 (dominant peptide 50)

[0985] AGAAVKGIGTM

[0986] SEQ ID NO: 78 (dominant peptide 62) KGKFQTAAQRAMVDQ

[0987] SEQ ID NO: 79 (dominant peptide 86)

[0988] AHLSQLVWMA

[0989] SEQ ID NO: 80 (dominant peptide 110)

[0990] PFEKSTIMAAFTGNT

[0991] SEQ ID NO: 81 (dominant peptide 49)

[0992] RSGAAGAAVKGIGTM

[0993] SEQ ID NO: 82 (dominant peptide 61)

[0994] CNILKGKFQTAAQRA

[0995] SEQ ID NO: 83 (dominant peptide 85)

[0996] NENPAHKSQLVWMAC

[0997] It is understood that the foregoing detailed description and accompanying examples are merely illustrative and are not to be taken as limitations upon the scope of the invention, which is defined solely by the appended claims and their equivalents.

[0998] Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications, including without limitation those relating to the chemical structures, substituents, derivatives, intermediates, syntheses, compositions, formulations, or methods of use of the invention, may be made without departing from the spirit and scope thereof.

Claims

CLAIMSWhat is claimed is:

1. An anti -HA (influenza a virus hemagglutinin) antibody or fragment thereof.

2. The anti -HA antibody, or fragment thereof, of claim 1, comprising a light chain selected from the group consisting of: a) SEQ ID NO:26, SEQ ID NO:

28. SEQ ID NO:30, SEQ ID NO: 32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, and SEQ ID NO:46; b) an amino acid sequence at least 90% identical to SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO 44, and SEQ ID NO:46; c) an amino acid sequence at least 70% of the length of SEQ ID NO:

26. SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:

36. SEQ ID NO:38, SEQ ID NO:

40. SEQ ID NO:42, SEQ ID NO:44, and SEQ ID NO:46; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:26, SEQ ID NO:

28. SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO 40, SEQ ID NO:42, SEQ ID NO:44, and SEQ ID NO:46.

3. The anti-HA antibody, or fragment thereof, of claim 1 or claim 2, comprising a heavy chain selected from the group consisting of: a) SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NOTO, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO 20, SEQ ID NO:22, and SEQ ID NO:24; b) an amino acid sequence at least 90% identical to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:

8. SEQ ID NOTO, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO:16, SEQ ID NO: 18, SEQ ID NO 20, SEQ ID NO:22, and SEQ ID NO:24; c) an amino acid sequence at least 70% of the length of SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:

8. SEQ ID NOTO, SEQ ID NO: 12, SEQ IDNO:

14. SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO 20, SEQ ID NO:22, and SEQ ID NO:24: and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO:20, SEQ ID NO:22, and SEQ ID NO:24.

4. The anti -HA antibody or fragment thereof of any one of claims 1-3, wherein the antibody is selected from the group consisting of a humanized antibody, a chimeric antibody, a fully human antibody, and an antibody mimetic.

5. A nucleic acid molecule encoding an anti-HA antibody, or fragment thereof, of any one of claims 1-4.

6. The nucleic acid molecule of claim 5, wherein the nucleotide sequence encoding the light chain of an anti-HA antibody is selected from the group consisting of: a) SEQ ID NO:25, SEQ ID NO:

27. SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO 39, SEQ ID NO:41 , SEQ ID NO:43, and SEQ ID NO:45; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:

37. SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO 43, and SEQ ID NO:45; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:

25. SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO 31, SEQ ID NO:33, SEQ ID NO:

35. SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, and SEQ ID NO:45; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, and SEQ ID NO:45.

7. The nucleic acid molecule of claim 5 or claim 6, wherein the nucleotide sequence encoding the heavy chain of an anti-HA antibody is selected from the group consisting of:a) SEQ ID NO:1, SEQ ID N0:3, SEQ ID N0:5, SEQ ID N0:7, SEQ ID N0:

9. SEQ ID NO: 11, SEQ ID N0: 13, SEQ 1D NO:

15. SEQ ID N0: 17, SEQ 1D NO:

19. SEQ ID N0:21, and SEQ ID NO:23; b) a nucleic acid sequence at least 90% identical to SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO:15, SEQ ID NO: 17, SEQ ID NO 19, SEQ ID NO:21, and SEQ ID NO:23; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ IDNO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, and SEQ ID NO:23; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO:

13. SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:

21. and SEQ ID NO:23.

8. An anti-NA (influenza a virus neuraminidase) antibody or fragment thereof.

9. The anti-NA antibody, or fragment thereof, of claim 8, comprising a light chain selected from the group consisting of: a) SEQ ID NO:49 and SEQ ID NO:53; b) an amino acid sequence at least 90% identical to SEQ ID NO:49 and SEQ ID NO:53; c) an amino acid sequence at least 70% of the length of SEQ ID NO:49 and SEQ ID NO:53; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:49 and SEQ ID NO:53.

10. The anti-NA antibody, or fragment thereof, of claim 8 or claim 9, comprising a heavy chain selected from the group consisting of: a) SEQ ID NO:47 and SEQ ID NO:51; b) an amino acid sequence at least 90% identical to SEQ ID NO: 47 and SEQ ID NO:51;114c) an amino acid sequence at least 70% of the length of SEQ ID NO:47 and SEQ ID NO:51; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:47 and SEQ ID NO:51.

11. The anti -NA antibody or fragment thereof of any one of claims 8-10, wherein the antibody is selected from the group consisting of a humanized antibody, a chimeric antibody, a fully human antibody, and an antibody mimetic.

12. A nucleic acid molecule encoding an anti-NA antibody, or fragment thereof, of any one of claims 8-11.

13. The nucleic acid molecule of claim 12, wherein the nucleotide sequence encoding the light chain of an anti-NA antibody is selected from the group consisting of: a) SEQ ID NO:50 and SEQ ID NO:54; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:50 and SEQ ID NO:54; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:50 and SEQ ID NO:54; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:50 and SEQ ID NO:54.

14. The nucleic acid molecule of claim 12 or claim 13, wherein the nucleotide sequence encoding the heavy chain of an anti-NA antibody is selected from the group consisting of: a) SEQ ID NO:48 and SEQ ID NO:52; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:48 and SEQ ID NO:52; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:48 and SEQ ID NO:52; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:48 and SEQ ID NO:52.

15. An influenza A nucleoprotein (NP) polypeptide.11516. The polypeptide of claim 15, comprising an amino acid sequence selected from the group consisting of: a) SEQ ID NO: 56 or SEQ ID NO: 58; b) an amino acid sequence at least 90% identical to SEQ ID NO: 56 or SEQ ID NO: 58; c) an amino acid sequence at least 70% of the length of SEQ ID NO:56 or SEQ ID NO: 58; and d) an amino acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 56 or SEQ ID NO: 58.

17. A nucleic acid molecule encoding a polypeptide of claim 15 or 16.

18. The nucleic acid molecule of claim 17. wherein the nucleotide sequence is selected from the group consisting of: a) SEQ ID NO:55 or SEQ ID NO:57; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:55 or SEQ ID NO:57; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:55 or SEQ ID NO:57; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:55 or SEQ ID NO:57.

19. An influenza A hemagglutinin (HA) protein polypeptide.

20. The polypeptide of claim 19, comprising an amino acid sequence selected from the group consisting of: a) SEQ ID NO: 60; b) an ammo acid sequence at least 90% identical to SEQ ID NO: 60; c) an amino acid sequence at least 70% of the length of SEQ ID NO: 60; and d) an ammo acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO: 60.11621. A nucleic acid molecule encoding a polypeptide of claim 19 or 20.

22. The nucleic acid molecule of claim 21, wherein the nucleotide sequence is selected from the group consisting of: a) SEQ ID NO:59; b) a nucleic acid sequence at least 90% identical to SEQ ID NO:59; c) a nucleic acid sequence at least 70% of the length of SEQ ID NO:59; and d) a nucleic acid sequence at least 90% identical to and at least 70% of the length of SEQ ID NO:59.

23. The nucleic acid molecule of any one of claims 5-7, 12-14, 17, 18, 21, and 22. wherein the nucleotide sequence encodes a leader sequence.

24. The nucleic acid molecule of any one of claims 5-7, 12-14, 17, 18, 21, and 22. wherein the nucleic acid molecule comprises an expression vector.

25. A composition comprising at least one nucleic acid molecule of any one of claims 5-7, 12-14, 17, 18, and 21-24.

26. A composition comprising: a) at least one anti-HA antibody or fragment thereof of any one of claims 1-4; b) at least one anti-NA antibody or fragment thereof of any one of claims 8-11 ; c) at least one polypeptide of claim 15 or 16; or d) at least one polypeptide of claim 19 or 20.

27. The composition of claim 25 or claim 26, further comprising a pharmaceutically acceptable excipient.

28. A method of preventing or treating a disease in a subject, the method comprising administering to the subject the antibody or antibody fragment of any one of claims 1-4 or 8-11, the polypeptide of claim 15, 16, 19, or 20, the nucleic acid molecule of117any one of claims 5-7, 12-14, 17, 18, and 21-24, a composition of any one of claims 25-27, or any combination thereof.

29. The method of claim 28, wherein the disease is influenza.118

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