Anti-cd73 antibodies and uses thereof

Abstract

Provided is an isolated antigen binding protein having one or more of the following properties: a. capable of binding to human CD73 and CD73 of cynomolgus monkey; b. capable of inhibiting 5' exonucleotidase activity of CD73; c. capable of modulating internalization of CD73; d. capable of promoting T cell proliferation; e. relatively stable in concentration in serum; and f. capable of inhibiting tumor growth and / or tumor cell proliferation. Furthermore, provided is a method for preparing the isolated antigen binding protein and a pharmaceutical use of the isolated antigen binding protein in preventing, alleviating and / or treating tumors.

Description

[0001] This application is a divisional application of Chinese patent application filed on August 20, 2020, with application number 2020800059738 and invention title "Anti-CD73 Antibody and Its Application". Background Technology

[0002] Leukocyte differentiation antigen 73, CD73, also known as 5'-nucleotidase, is an enzyme encoded by the NT5E gene in the human body. CD73 is a dimer composed of two identical 70-kD subunits, with a glycosylphosphatidylinositol atom attached to the outer surface of the plasma membrane.

[0003] CD73 dephosphorylates extracellular monophosphate (AMP) to produce adenosine. The accumulation of extracellular adenosine in cancerous tissue constitutes an important mechanism for tumor immune escape. Furthermore, tumor-derived adenosine significantly inhibits infiltrating effector T cells through A2A receptors activated by adenylate cyclase (Ohta et al., (2006) Proc Natl Acad Sci USA 103:13132-13137). Studies have shown that CD73 is highly expressed on the surface of various tumor cells, including leukemia, bladder cancer, glioma, glioblastoma, ovarian cancer, melanoma, prostate cancer, thyroid cancer, esophageal cancer, and breast cancer (Jin et al., Cancer Res 2010; 70:2245-55 and Stagg et al., PNAS 2010; 107:1547-52).

[0004] CD73 has been shown to regulate cell-cell and cell-matrix interactions on tumor cells, and its expression and activity are also associated with reduced T-cell responses and drug resistance. Another issue is that antibodies commonly referred to as CD73 inhibitors may not function by modulating the 5' exonuclease activity of CD73. One antibody, 7G2 (mlgG2 isotype, Life Technologies), has been reported to inhibit CD73, but this antibody does not bind to cell-surface CD73 in flow cytometry, or at most binds with very low affinity. Another CD73-binding antibody, clone AD2 (mouse IgG1 isotype), has been reported to induce receptor aggregation and internalization, but has minimal effect on enzyme activity. However, another reagent, 1E9 (mouse IgG3 isotype, Santa Cruz Biotechnology Ltd.), can promote T-cell signaling transduction independently of enzymatic inhibition. Another mAb, 4G4 (IgG1 isotype, Novus Biologicals), has been reported to induce CD73 detachment from the T-cell surface. In experiments using recombinant CD73, only one reagent, although not further characterized, has been reported to have the ability to partially block enzymatic activity (Sachsenmeier et al. (2012) J. Biomed. Screening 17:993-998), which was later defined as an antibody capable of inducing intracellularization (Rust et al., (2013) Mol. Cancer 12:11). Furthermore, a more complicating factor is that the antibodies described in the literature are often murine isotypes capable of binding to the Fcγ receptor, making it difficult to separate any potential blocking effect from Fc-mediated action. Anti-CD73 antibodies that bind to the Fcγ receptor may, for example, modulate the depletion of CD73-expressing tumor cells (and possibly CD73-expressing immunosuppressive cells) (e.g., through ADCC), and / or induce the production of inflammatory cytokines, rather than any true blocking effect. Therefore, despite considerable interest in research targeting CD73, the characterization of most effective anti-CD73 antibodies remains to be determined.

[0005] New diagnostic methods and antibodies targeting CD73 need to be developed to realize its potential as a novel therapeutic target. Summary of the Invention

[0006] This application relates to an isolated antigen-binding protein comprising one or more of the following properties: a. capable of binding to human CD73 and cynomolgus monkey CD73 with a considerably high affinity; b. capable of inhibiting the 5′ exonuclease activity of CD73; c. capable of regulating CD73 internalization; d. capable of relieving the inhibitory effect of adenosine on T cell proliferation; e. relatively stable concentration in serum; and f. capable of inhibiting tumor growth and / or tumor cell proliferation. Furthermore, this application also provides a method for preparing the aforementioned antigen-binding protein and the pharmaceutical use of the said antigen-binding protein in the prevention, mitigation, and / or treatment of tumors.

[0007] On the one hand, this application provides an isolated antigen-binding protein, including one or more of the following properties: a. capable of simultaneously binding human CD73 and cynomolgus monkey CD73 with a considerably high affinity; b. capable of inhibiting the 5′ exonuclease activity of CD73; c. capable of regulating the internalization of CD73; d. capable of relieving the inhibitory effect of adenosine on T cell proliferation; e. maintaining a relatively stable concentration in serum for at least 15 days; and f. capable of inhibiting tumor growth and / or tumor cell proliferation.

[0008] In some embodiments, the isolated antigen-binding protein includes any one of the HCDR1, HCDR2, and / or HCDR3 regions of the variable region VH of the antibody heavy chain shown in SEQ ID NO:168.

[0009] In some embodiments, the isolated antigen-binding protein includes any one of the LCDR1, LCDR2, and / or LCDR3 regions of the variable region VL of the antibody light chain shown in SEQ ID NO:169.

[0010] In some embodiments, the isolated antigen-binding protein includes an antibody or an antigen-binding fragment.

[0011] In some embodiments, the antigen-binding fragment includes Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di-scFv and / or dAb.

[0012] In some embodiments, the antibody is selected from monoclonal antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.

[0013] In some embodiments, the antigen-binding protein comprises any one of the HCDR1, HCDR2, and / or HCDR3 regions from the heavy chain variable region VH shown in SEQ ID NO:152.

[0014] In some embodiments, the VH comprises an amino acid sequence shown in any one of SEQ ID NO: 96, 104, 105, 106 and 107.

[0015] In some embodiments, the isolated antigen-binding protein comprises any one of the LCDR1, LCDR2, and / or LCDR3 regions of the light chain variable region VL shown in SEQ ID NO:151.

[0016] In some embodiments, the VL comprises the amino acid sequence shown in either SEQ ID NO:111 or 115.

[0017] In some embodiments, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:154.

[0018] In some embodiments, the HCDR3 comprises any one of the amino acid sequences shown in SEQ ID NO:35, 38, and 39.

[0019] In some embodiments, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:153.

[0020] In some embodiments, the HCDR2 comprises the amino acid sequence shown in any one of SEQ ID NO:18,23 and24.

[0021] In some embodiments, the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:7.

[0022] In some embodiments, the isolated antigen-binding protein includes a heavy chain variable region VH, wherein the VH contains the amino acid sequence shown in SEQ ID NO:152.

[0023] In some embodiments, the VH comprises the amino acid sequence shown in any one of SEQ ID 96, 104, 105, 106 and 107.

[0024] In some embodiments, the isolated antigen-binding protein comprises an antibody heavy chain HC, said HC comprising an amino acid sequence represented by any one of SEQ ID NO: 124, 132, 133, 134, and 135.

[0025] In some embodiments, the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:78.

[0026] In some embodiments, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:66.

[0027] In some embodiments, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:55.

[0028] In some embodiments, the isolated antigen-binding protein comprises a light chain variable region VL, wherein the VL comprises the amino acid sequence shown in SEQ ID NO:151.

[0029] In some embodiments, the VL comprises the amino acid sequence shown in either SEQ ID NO:111 or 115.

[0030] In some embodiments, the isolated antigen-binding protein comprises an antibody light chain LC, the LC comprising the amino acid sequence shown in either SEQ ID NO: 139 or 143.

[0031] In some embodiments, the isolated antigen-binding protein comprises any one of the HCDR1, HCDR2, and / or HCDR3 regions from the heavy chain variable region VH shown in SEQ ID NO:156.

[0032] In some embodiments, the VH comprises an amino acid sequence shown in any one of SEQ ID NO: 95, 99, 100, 101, 102, and 103.

[0033] In some embodiments, the isolated antigen-binding protein comprises any one of the LCDR1, LCDR2, and / or LCDR3 regions of the light chain variable region VL shown in SEQ ID NO:155.

[0034] In some embodiments, the VL comprises an amino acid sequence represented by any one of SEQ ID NO: 110, 114, 118, 119, 120, 121, and 122.

[0035] In some embodiments, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:34.

[0036] In some embodiments, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:158.

[0037] In some embodiments, the HCDR2 is selected from amino acid sequences comprising any one of SEQ ID NO: 17, 21 and 22.

[0038] In some embodiments, the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:6.

[0039] In some embodiments, the isolated antigen-binding protein includes a heavy chain variable region VH, wherein the VH contains the amino acid sequence shown in SEQ ID NO:156.

[0040] In some embodiments, the VH comprises an amino acid sequence shown in any one of SEQ ID NO: 95, 99, 100, 101, 102 and 103.

[0041] In some embodiments, the antigen-binding protein comprises an antibody heavy chain HC, wherein the HC comprises an amino acid sequence shown in any one of SEQ ID NO: 123, 127, 128, 129, 130 and 131.

[0042] In some embodiments, the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:157.

[0043] In some embodiments, the LCDR3 comprises the amino acid sequence shown in any one of SEQ ID NO:77,81,84,85,86,87 and88.

[0044] In some embodiments, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:65.

[0045] In some embodiments, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:54.

[0046] In some embodiments, the isolated antigen-binding protein comprises a light chain variable region VL, wherein the VL comprises the amino acid sequence shown in SEQ ID NO:155.

[0047] In some embodiments, the VL comprises an amino acid sequence represented by any one of SEQ ID Nos: 110, 114, 118, 119, 120, 121, and 122.

[0048] In some embodiments, the isolated antigen-binding protein comprises an antibody light chain LC, wherein the LC comprises the amino acid sequence shown in any one of SEQ ID NO: 138, 142, 146, 147, 148, 149 and 150.

[0049] In some embodiments, the isolated antigen-binding protein comprises any one of the HCDR1, HCDR2, and / or HCDR3 regions from the heavy chain variable region VH shown in SEQ ID NO:108.

[0050] In some embodiments, the isolated antigen-binding protein comprises any one of the LCDR1, LCDR2, and / or LCDR3 regions from the light chain variable region VL shown in SEQ ID NO:116.

[0051] In some embodiments, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:40.

[0052] In some embodiments, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:25.

[0053] In some embodiments, the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:10.

[0054] In some embodiments, the isolated antigen-binding protein comprises a heavy chain variable region VH, the VH comprising the amino acid sequence shown in SEQ ID NO: 108.

[0055] In some embodiments, the isolated antigen-binding protein comprises an antibody heavy chain HC, the HC comprising the amino acid sequence shown in SEQ ID NO:136.

[0056] In some embodiments, the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:82.

[0057] In some embodiments, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:69.

[0058] In some embodiments, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:58.

[0059] In some embodiments, the isolated antigen-binding protein comprises a light chain variable region VL, the VL comprising the amino acid sequence shown in SEQ ID NO:116.

[0060] In some embodiments, the isolated antigen-binding protein comprises an antibody light chain LC, the LC comprising the amino acid sequence shown in SEQ ID NO:144.

[0061] On the other hand, this application provides one or more isolated nucleic acid molecules encoding the isolated antigen-binding protein.

[0062] On the other hand, this application provides one or more carriers that can contain the one or more isolated nucleic acid molecules.

[0063] On the other hand, this application provides cells comprising one or more isolated nucleic acid molecules and one or more carriers.

[0064] On the other hand, this application provides a method for preparing the isolated antigen-binding protein, comprising culturing the cells under conditions that cause the isolated antigen-binding protein to be expressed.

[0065] On the other hand, this application provides a pharmaceutical composition comprising an isolated antigen-binding protein, one or more isolated nucleic acid molecules, one or more carriers, cells, and optionally a pharmaceutically acceptable carrier.

[0066] On the other hand, this application provides the use of the isolated antigen-binding protein, the one or more nucleic acid molecules, the one or more carriers, the cells, and / or the pharmaceutical composition in the preparation of a medicament for the prevention, relief, and / or treatment of tumors.

[0067] In some embodiments, the tumor includes solid tumors and / or hematologic malignancies.

[0068] On the other hand, this application provides methods for preventing, alleviating and / or treating tumors, the methods comprising administering the isolated antigen-binding protein of this application to a subject in need.

[0069] In some embodiments, the tumor includes solid tumors and / or hematologic malignancies.

[0070] On the other hand, this application provides a method for inhibiting the 5'-nucleotidase activity of CD73, including the administration of the isolated antigen-binding protein of this application.

[0071] On the other hand, this application provides a method for regulating CD73 internalization, including the administration of the isolated antigen-binding protein of this application.

[0072] On the other hand, this application provides the use of the isolated antigen-binding protein, the one or more nucleic acid molecules, the one or more carriers, the cells and / or the pharmaceutical composition to prepare a medicament for the prevention, relief and / or treatment of tumors.

[0073] In some embodiments, the tumor includes solid tumors and / or hematologic malignancies.

[0074] On the other hand, this application provides a method for preventing, alleviating and / or treating tumors, including administering the isolated antigen-binding protein of this application to a subject in need.

[0075] Other aspects and advantages of this application will readily apparent to those skilled in the art from the detailed description below. The following detailed description shows and describes only exemplary embodiments of this application. It will be appreciated that this application can have other different embodiments, and that several details thereof can be modified in various obvious ways without departing from this disclosure. Accordingly, the drawings and description should be considered illustrative in nature, not restrictive.

[0076] Reference Merging

[0077] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference to the same extent as each individual publication, patent or patent application is expressly and individually indicated by reference. Attached Figure Description

[0078] The novel features of the present invention are specifically set forth in the appended claims. The features and advantages of the invention can be better understood by referring to the following detailed description, which illustrates illustrative embodiments employing the principles of the invention, along with the accompanying drawings, wherein:

[0079] Figure 1A and 1B The image shows the binding of the flow cytometry (FACS) antibody to the CHO-K1-hCD73 cell line.

[0080] Figure 2 The image shows the binding of a flow cytometry (FACS) antibody to the HEK293-cynoCD73 cell line.

[0081] Figure 3 The image shows the binding of a flow cytometry (FACS) antibody to the CHO-K1-hCD73 cell line.

[0082] Figure 4 The image shows the binding of the flow cytometry (FACS) antibody to the CHO-K1-hCD73 cell line.

[0083] Figure 5A and 5B The image shows the binding of a flow cytometry (FACS) antibody to the CHO-K1-hCD73 cell line.

[0084] Figure 6A and 6B The image shows antibody inhibition of cell-based CD73 enzyme activity.

[0085] Figure 7A Figures 7B, 8A, and 8B show the inhibition of soluble CD73 enzyme activity by antibodies treated at different time points.

[0086] Figure 9A Figures 9B, 10, and 11 show the internalization activity of the antibody.

[0087] Figure 12A Figures 12B, 13A, and 13B show the results of antibody assays in primitive T cell proliferation.

[0088] Figure 14AFigures 14B and 14C show the serum concentration-time curves (PK) results for each antibody: A: PR000506, B: PR000846, and C: PR000497.

[0089] Figure 15A Figures 15B, 16A, and 16B show the antibody's activity in inhibiting tumor growth in vivo, where A represents tumor volume and B represents body weight.

[0090] Figure 17 The diagram shows the difference in deuterium uptake between the CD73+PR000846 complex (positive value on the y-axis) and CD73 alone (negative value on the y-axis).

[0091] Figure 18 Figures 19 and 20 show the relative intake of deuterium (Dalton mass change) as a function of deuterium exposure time in the peptides containing regions 1, 2, and 3.

[0092] Figure 21 and Figure 22 The results shown are epitope analysis results of the combination of CD73 and PR000846. Detailed description

[0093] Although various embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, modifications, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.

[0094] Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the” used in this article generally include the plural forms.

[0095] As used herein, the term "CD73," also known as a 5′ exonuclease, generally refers to an enzyme (nuclease) capable of converting extracellular nucleotide 5′ monophosphate into nucleosides, specifically adenosine monophosphate (AMP) into adenosine. The primary function of CD73 is to convert extracellular nucleotides (e.g., 5′-AMP) into the highly immunosuppressive molecule adenosine. The term "CD73" includes any variant or isoform of CD73 naturally expressed in cells. CD73 or any variants and isoforms thereof can be isolated from cells or tissues that naturally express them, or can be recombinantly generated using techniques well known in the art and / or those described herein. Accordingly, the antigen-binding proteins described herein can cross-react with species other than humans (e.g., cynomolgus monkey CD73). Alternatively, antibodies can be specific to human CD73 and may not exhibit any cross-reactivity with other species. The amino acid sequence of human CD73 is shown in GenBank as accession number AAH65937.1 (5'-nucleotidase, cell membrane), NP_002517 (isoform 1 preprotein) or NP_001191742 (isoform 2 preprotein).

[0096] As used herein, the term “Tab1” or PR002078 generally refers to an anti-CD73 antibody. The heavy chain amino acid sequence of Tab1 is shown in SEQ ID NO.137, and the light chain amino acid sequence of Tab1 is shown in SEQ ID NO.145.

[0097] As used herein, the term “Tab2” or PR000752 generally refers to an anti-CD73 antibody. The heavy chain amino acid sequence of Tab2 is shown in SEQ ID NO.126, and the light chain amino acid sequence of Tab2 is shown in SEQ ID NO.141.

[0098] As used herein, the term “Tab3” or PR000690 generally refers to an anti-CD73 antibody. The heavy chain amino acid sequence of Tab3 is shown in SEQ ID NO.125, and the light chain amino acid sequence of Tab2 is shown in SEQ ID NO.140.

[0099] As used herein, the term “internalization” refers to the molecular, biochemical, and cellular events associated with the process by which molecules are transferred from the extracellular surface of a cell to the intracellular surface. This process can involve, in particular, the internalization of extracellular molecules (e.g., hormones, antibodies, and small organic molecules), membrane-associated molecules (e.g., cell surface receptors), and complexes of membrane-associated molecules that bind to extracellular molecules (e.g., ligands that bind to transmembrane receptors or antibodies that bind to membrane-associated molecules).

[0100] As used herein, the term "antigen-binding protein" generally refers to a protein comprising an antigen-binding portion and optionally a scaffold or framework portion that allows the antigen-binding portion to acquire a conformation that promotes antigen-binding. Examples of antigen-binding proteins include antibodies, antibody fragments (e.g., the antigen-binding portion of an antibody), antibody derivatives, and antibody analogs. Antigen-binding proteins can have, for example, the structure of naturally occurring immunoglobulins. An "immunoglobulin" is a tetrameric molecule. In naturally occurring immunoglobulins, each tetramer consists of two pairs of identical polypeptide chains, each pair having one light chain (LC) and one heavy chain (HC). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids, primarily responsible for antigen recognition. The carboxyl-terminal portion of each chain defines a constant region, primarily responsible for effector function. Human light chains are classified as κ or λ light chains. Heavy chains are classified as μ, δ, γ, α, or ε, and antibody isotypes are defined as IgM, IgD, IgG, IgA, and IgE, respectively. The variable regions of naturally occurring immunoglobulin chains generally exhibit a structure of four relatively conserved framework regions (FRs) connected by three highly variable regions (also called complementarity-determining regions or CDRs). From the N-terminus to the C-terminus, both the light and heavy chains contain the domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0101] As used herein, the term "CDR" generally refers to the complementarity-determining region (CDR) amino acid sequence of an antigen-binding protein, which is a hypervariable region of the heavy and light chains of an immunoglobulin. There are three heavy chain and three light chain CDRs (or CDR regions) in the variable portion of an immunoglobulin. It will be apparent to those skilled in the art that various numbering systems exist for CDR sequences, such as Chothia (Chothia et al. (1989) Nature 342: 877-883) and Kabat (Kabat et al., Protein Sequences in Immunology, National Institutes of Health, Bethesda, Maryland (1987 and 1991)). The Chothia numbering system is used to number residues in the antibodies disclosed herein.

[0102] Kabat Chothia Combined LCDR1 L24-L34 L24-L34 L24-L34 LCDR2 L50-L56 L50-L56 L50-L56 LCDR3 L89-L97 L89-L97 L89-L97 HCDR1 H31-H35 H26-H32 H26-H35 HCDR2 H50-H65 H52-H56 H50-H65 HCDR3 H95-H102 H95-H102 H95-H102

[0103] Among them, Laa-Lbb can refer to the amino acid sequence from position aa to position bb starting from the N-terminus of the antibody; for example, L24-L34 can refer to the amino acid sequence from position 24 to position 34 of the antibody light chain;

[0104] The term “isolated” when used for nucleic acids or proteins means that the nucleic acid or protein is essentially free of other cellular components associated with it in its natural state.

[0105] As used in this article, the term "K" D"Affinity" or "KD" usually refers to the dissociation constant, a specific type of equilibrium constant used to measure the tendency of a larger object to reversibly separate (dissociate) into smaller components, as when a complex breaks down into its component molecules. The dissociation constant is the reciprocal of the association constant. In the specific case of antibody binding to an antigen, the term affinity constant usually refers to the association constant.

[0106] As used herein, the term "antigen-binding protein" generally refers to a portion of an immunoglobulin molecule. An antigen-binding fragment may consist of a light chain and a portion of a heavy chain having a single antigen-binding site. Antigen-binding fragments can be obtained by digesting immunoglobulin molecules with papain. For example, an antigen-binding fragment may consist of a constant domain and a variable domain, each in the heavy and light chains. The variable domain may contain a complementary site (antigen-binding site) at the amino terminus of the immunoglobulin molecule, which includes a set of complementarity-determining regions. Papain can be used to cleave immunoglobulin molecules into two Fab fragments and one Fc fragment. Pepsin cleaves below the hinge region, thus forming the F(ab')2 fragment and the pFc' fragment. The divalent F(ab)2 or F(ab')2 fragment has two antigen-binding regions linked by disulfide bonds. Reduction of the F(ab)2 or F(ab')2 fragment yields two monovalent Fab or Fab' fragments, which have free thiol groups that can bind to other molecules.

[0107] As used in this article, the term "Fv fragment" generally refers to the smallest fragment produced by enzymatic cleavage of IgG and IgM antibodies.

[0108] As used herein, the term "ScFv" typically refers to a single-chain antibody fragment. ScFv can be a recombinant single-chain polypeptide molecule in which the variable regions of the light and heavy chains of the antibody are linked directly or via peptide linkers.

[0109] As used herein, the term “fully humanized antibody” generally refers to an antibody having antibody regions derived entirely from human amino acid sequences, wherein antigen-specific selection is performed in vivo using genetically modified mice or through antibody engineering methods that combine screening.

[0110] As used herein, the term "one or more isolated nucleic acid molecules" generally refers to a polymeric form of nucleotides of any length isolated from their natural environment or synthesized artificially, said nucleotides being deoxyribonucleotides or ribonucleotides or analogues thereof.

[0111] As used herein, the term "one or more vectors" generally refers to a nucleic acid medium that allows the insertion and expression of polynucleotides encoding proteins. Genetic material elements carried in a vector can be expressed in a host cell by transformation, transduction, or transfection with the vector. Examples of vectors include plasmids, phage particles, granules, artificial chromosomes (e.g., yeast artificial chromosome (YAC), bacterial artificial chromosome (BAC), or P1-derived artificial chromosome (PAC)), bacteriophages (e.g., λ phage or M13 phage), and animal viruses. Vectors may contain a variety of elements that control expression, including promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. Additionally, vectors may contain an origin of replication. Vectors may also contain components that facilitate their entry into the cell, such as viral particles, liposomes, or protein shells, but are not limited to these substances.

[0112] As used herein, the term "cell" generally refers to a cell that can be used to carry one or more vectors of this application, or to express or produce antibodies, antigen-binding fragments, or variants thereof of this application. The cell in this application can be a host cell. This cell can be a prokaryotic cell, such as *Escherichia coli* and *Bacillus subtilis*; or a fungal cell, such as yeast cells or *Aspergillus* cells; an insect cell, such as S2 *Drosophila* cells or Sf9 cells; or another cell type, such as CHO cells, COS cells, NSO cells, or other cells suitable for antibody expression.

[0113] As used herein, the term "conditions for expression" generally refers to conditions that enable the expression of the isolated antigen-binding proteins of this disclosure. In some embodiments, such conditions may include, but are not limited to, incubation time, temperature, and culture medium, and may depend on cell type and can be readily determined by a skilled technician.

[0114] Isolated antigen-binding proteins

[0115] In one aspect, this disclosure provides isolated antigen-binding proteins capable of binding to human CD73 and cynomolgus monkey CD73 with comparable binding affinity. As used herein, the term "comparable binding affinity" generally refers to a ratio of the two binding affinities of the antibody from about 0.01 nM to about 100 nM, for example, from about 0.02 nM to about 50 nM, from about 0.05 nM to about 30 nM, from about 0.1 nM to about 20 nM, from about 0.1 nM to about 10 nM, from about 0.2 nM to about 10 nM, and from about 0.5 nM to about 3 nM. The binding affinity of the antibody to CD73 can be assessed using CD73 stable cell lines (e.g., the CHO-K1 cell line), for example, in FACS analysis.

[0116] The isolated antigen-binding protein inhibits the 5′ exonuclease activity of CD73. The isolated antigen-binding protein inhibits CD73-mediated adenosine production, specifically inhibiting CD73-mediated AMP catabolism to adenosine. This can be assessed by testing the ability of the antibody to directly or indirectly block the conversion of AMP to adenosine. In some cases, the isolated antigen-binding protein can inhibit cell-based CD73 activity. This can be measured by constructing a cell line stably expressing CD73, such as the CHO-K1 cell line. For example, the isolated antigen-binding protein can inhibit 50% of the 5′ exonuclease activity of cell-based CD73 at concentrations as described in the examples, from about 0.01 nM to about 100 nM, for example, from about 0.02 nM to about 50 nM, from about 0.001 nM to about 10 nM, from about 0.001 nM to about 1 nM, from about 0.01 nM to about 0.5 nM, from about 0.01 nM to about 0.3 nM, and from about 0.05 nM to about 0.25 nM. In another case, the isolated antigen-binding protein can inhibit the activity of soluble recombinant CD73. For example, by quantifying the enzymatic product inorganic phosphate ester, the isolated antigen-binding protein can cause a reduction in the conversion of AMP to adenosine by at least 40%, at least 50%, at least 60%, at least 70%, or at least 90%.

[0117] Isolated antigen-binding proteins may be able to mediate CD73 internalization. In some cases, the percentage of internalization can be assessed by measuring the residual CD73 on the cell surface, such as NCI-H292, using flow cytometry. For example, after 4 hours of incubation with an antibody, approximately 40% to approximately 99% (e.g., approximately 40% to approximately 90%, approximately 40% to approximately 80%, approximately 40% to approximately 70%, approximately 40% to approximately 60%, approximately 40% to approximately 55%, or approximately 50%) of CD73 is internalized into the cell. In another case, internalization can be measured using an internalization kit as described in the examples. After 3 days of incubation with an antibody, CD73 on the surface of CHO-K1 is internalized, and the maximum level of antibody-mediated CD73 internalization may be at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more, depending on the antibody concentration.

[0118] The activity of isolated antigen-binding proteins can also be measured in indirect assays of their ability to regulate lymphocyte activity, such as the ability to alleviate AMP-mediated lymphocyte activity inhibition or to induce lymphocyte proliferation. In some cases, the ability of isolated antigen-binding proteins to regulate lymphocyte proliferation can be assessed by indirect assays. Lymphocytes can be T cells, including helper T cells (e.g., CD4+ T cells). For example, FACS can be used for analysis.

[0119] The isolated antigen-binding protein can maintain a relatively stable concentration in serum for at least 15 days. As used herein, "relatively stable" generally means that the concentration of the antibody in serum does not change by more than 50% compared to a reference concentration (e.g., the initial concentration of the antibody in serum). The stability of the antibody in serum can be determined by pharmacokinetic analysis. For example, as described in the PK analysis, the t-strength of the isolated antigen-binding protein of this disclosure is... 1 / 2 It takes about 15 days.

[0120] In this application, the antigen-binding protein may include at least one HCDR from VH of SEQ ID NO: 152.

[0121] In this application, the antigen-binding protein may comprise HCDR3 from the heavy chain variable region VH of SEQ ID NO: 152, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 154: X1X2QWGSRLDY (SEQ ID NO. 154); wherein X1 = D or E; X2 = G or A. For example, the sequence may be determined according to the definition of Chothia.

[0122] In some cases, compared to HCDR3 of the antigen-binding protein DGQWGSRLDY, HCDR3 may contain at least one amino acid substitution selected from the following: amino acid substitutions at X1 and X2, wherein the amino acid at X1 may be substituted with E and the amino acid at X2 may be substituted with A.

[0123] For example, HCDR3 may contain an amino acid sequence as shown in any of SEQ ID NO: 35, 38 and 39.

[0124] In this application, the antigen-binding protein may comprise HCDR2 from the heavy chain variable region VH of SEQ ID NO: 152, and the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 153: WYX3X4SF (SEQ ID NO. 153); wherein X3 = D or E; X4 = G or A. For example, the sequence may be determined according to the definition of Chothia.

[0125] For example, HCDR2 may contain an amino acid sequence as shown in any of SEQ ID NO: 18, 23 and 24.

[0126] In this application, the antigen-binding protein may comprise HCDR1 from the heavy chain variable region VH of SEQ ID NO: 152, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: GFTFSDY (SEQ ID NO. 7). For example, the sequence can be determined according to the definition of Chothia.

[0127] In this application, the antigen-binding protein may include H-FR1, the C-terminus of which is directly or indirectly linked to the N-terminus of HCDR1, and H-FR1 may contain an amino acid sequence as shown in SEQ ID NO.2: QVQLVESGGGVAQPGRSLSLSCAAS (SEQ ID NO.2). For example, the sequence can be determined according to the definition of Chothia.

[0128] In this application, the antigen-binding protein may include H-FR2, which is located between HCDR1 and HCDR2, and H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12: GMHWVRQAPGKGLEWVALIWVRQAPGKGLEWVA (SEQ ID NO.12). For example, the sequence may be determined according to the definition of Chothia.

[0129] In this application, the antigen-binding protein may include H-FR3, which is located between HCDR2 and HCDR3, and H-FR3 may contain the amino acid sequence shown in SEQ ID NO: 28: EYYADSVKGRFSISRDNSKNTLYLQMNSLRAEDTAVYYCVR (SEQ ID NO: 28). For example, the sequence may be determined according to the definition of Chothia.

[0130] In this disclosure, the antigen-binding protein may comprise H-FR4, the N-terminus of which is directly or indirectly linked to the C-terminus of HCDR3, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.43: WGQGTLVTVSS (SEQ ID NO.43). For example, the sequence may be determined according to the definition of Chothia.

[0131] In this disclosure, the antigen-binding protein may include VH, and VH may include SEQ ID NO.152: QVQLVESGGGVAQPGRSLSLSCAASGFTFSDYGMHWVRQAPGKGLEWVALIWYX 54 X 55 SF EYYADSVKGRFSISRDNSKNTLYLQMNSLRAEDTAVYYCVRX 99 X 100 The amino acid sequence shown is QWGSRLDYWGQGT LVTVSS (SEQ ID NO.152); where X 54 =D or E; X 55 =G or A; X 99 = D or E; X100 =G or A. For example, a sequence can be determined according to the definition of Chothia.

[0132] For example, VH may contain an amino acid sequence as shown in any one of SEQ ID NO: 96, 104, 105, 106 and 107.

[0133] In this disclosure, the antigen-binding protein may include an antibody heavy chain constant region, and the antibody heavy chain constant region includes a human IgG constant region. In some cases, the human IgG constant region may be derived from the human IgG1 constant region. For example, the constant region of the antibody heavy chain can contain the amino acid sequence shown in SEQ ID NO.167: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO.167).

[0134] In this application, the antigen-binding protein may comprise an antibody heavy chain (HC), and the HC comprises an amino acid sequence as shown in SEQ ID NO. 162. For example, the HC may comprise an amino acid sequence as shown in any one of SEQ ID NO: 124, 132, 133, 134, and 135.

[0135] In this application, the antigen-binding protein may include at least one LCDR from VL of SEQ ID NO: 151.

[0136] In this application, the antigen-binding protein may comprise LCDR3 from the heavy chain variable region VH of SEQ ID NO: 152, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 78: QQRSNWIFT (SEQ ID NO. 78). For example, the sequence may be determined according to the definition of Chothia.

[0137] In this disclosure, the antigen-binding protein may comprise LCDR2 from the heavy chain variable region VH of SEQ ID NO. 152, and LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 66: DASNRAT (SEQ ID NO. 66). For example, the sequence may be determined according to the definition of Chothia.

[0138] In this application, the antigen-binding protein may comprise LCDR1 from the heavy chain variable region VH of SEQ ID NO. 152, and LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 55: RASQSVSRYLA (SEQ ID NO. 55). For example, the sequence may be determined according to the definition of Chothia.

[0139] In this application, the antigen-binding protein may comprise L-FR1, the C-terminus of which is directly or indirectly linked to the N-terminus of LCDR1, and L-FR1 may comprise the amino acid sequence shown in SEQ ID NO.48: X1IVX4TQSPATLSLSPGERATLSC (SEQ ID NO.48); wherein X1 = K or E; X4 = M or L. For example, the sequence can be determined according to the definition of Chothia.

[0140] In some cases, compared to L-FR1 of the antigen-binding protein shown in SEQ ID NO.47, L-FR1 may contain at least one amino acid substitution selected from X1 and / or X4, wherein the amino acid on X1 may be substituted with E and the amino acid on X4 may be substituted with L.

[0141] For example, L-FR1 may contain an amino acid sequence as shown in either SEQ ID NO: 48 or 51.

[0142] In this application, the antigen-binding protein may include L-FR2, which is located between LCDR1 and LCDR2, and L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61: WYQQKPGQAPRLLIY (SEQ ID NO.61). For example, the sequence may be determined according to the definition of Chothia.

[0143] In this application, the antigen-binding protein may include L-FR3, which is located between LCDR2 and LCDR3, and L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72: GIPARFSGSGSGTGTTLTISSLEPEDFAVYYC (SEQ ID NO.72). For example, the sequence may be determined according to the definition of Chothia.

[0144] In this application, the antigen-binding protein may comprise L-FR4, the N-terminus of which is directly or indirectly linked to the C-terminus of LCDR3, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO.90: FGPGTKVDIK (SEQ ID NO.90). For example, the sequence may be determined according to the definition of Chothia.

[0145] In this disclosure, the antigen-binding protein may comprise a VL, and the VL may comprise an amino acid sequence as shown in SEQ ID NO. 151: X1IVX4TQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPAR FSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWIFTFGPGTKVDIK (SEQ ID NO. 151), wherein X1 = K or E; X4 = M or L. For example, the sequence can be determined according to the definition of Chothia.

[0146] For example, VL may contain an amino acid sequence as shown in either SEQ ID NO: 111 or 115.

[0147] In this application, the antigen-binding protein may include an antibody light chain constant region, and the antibody light chain constant region may include a human Igκ constant region. For example, the antibody light chain constant region may include the amino acid sequence shown in SEQ ID NO: 166: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 166).

[0148] In this application, the antigen-binding protein may comprise an antibody light chain (LC), and the LC comprises an amino acid sequence as shown in SEQ ID NO. 159. For example, the LC may comprise an amino acid sequence as shown in either SEQ ID NO: 139 or 143.

[0149] In this application, the antigen-binding protein may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO.7, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO.153, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO.154.

[0150] In some embodiments, the antigen-binding protein may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO.7, HCDR2 may comprise the amino acid sequence shown in any one of SEQ ID NO.18, 23, and 24, and HCDR3 may comprise the amino acid sequence shown in any one of SEQ ID NO.35, 38, and 39.

[0151] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.7, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.18, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.35.

[0152] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.7, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.23, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.38.

[0153] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.7, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.23, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.39.

[0154] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.7, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.24, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.38.

[0155] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.7, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.24, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.39.

[0156] In this application, the antigen-binding protein may comprise LCDR1-3. LCDR1 may comprise the amino acid sequence shown in SEQ ID NO.55, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO.78.

[0157] In this application, the antigen-binding protein may comprise H-FR1-4. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO.2, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO.12, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.43.

[0158] In this application, the antigen-binding protein may comprise L-FR1-4. L-FR1 may comprise the amino acid sequence shown in SEQ ID NO.48, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO.61, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO.90.

[0159] In some cases, L-FR1 may contain the amino acid sequence shown in SEQ ID NO: 51, L-FR2 may contain the amino acid sequence shown in SEQ ID NO: 61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO: 72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO: 90.

[0160] In this application, the antigen-binding protein may comprise HCDR1-3 and LCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 7, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 153, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 154; LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 55, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 66, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 78.

[0161] In some cases, the HCDR1 of the antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO.7, the HCDR2 may contain the amino acid sequences shown in any one of SEQ ID NO.18, 23 and 24, and the HCDR3 may contain the amino acid sequence shown in SEQ ID NO.35; the LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, the LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and the LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78.

[0162] For example, the HCDR1 of the antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO.7, the HCDR2 may contain the amino acid sequence shown in SEQ ID NO.18, and the HCDR3 may contain the amino acid sequence shown in SEQ ID NO.35; the LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, the LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and the LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78.

[0163] For example, the antigen-binding protein HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.23, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.38; LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78.

[0164] In this application, the antigen-binding protein may comprise H-FR1-4 and L-FR1-4. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 2, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 12, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 28, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 43. L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 48, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 61, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 72, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 90.

[0165] In some cases, H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.51, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90.

[0166] In this application, the antigen-binding protein may comprise VH and VL. VH may comprise the amino acid sequence shown in SEQ ID NO: 152, and VL may comprise the amino acid sequence shown in SEQ ID NO: 151.

[0167] In some cases, the VH may contain an amino acid sequence as shown in any one of SEQ ID NO: 96, 104, 105, 106 and 107, and the VL may contain an amino acid sequence as shown in any one of SEQ ID NO: 111 and 115.

[0168] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 96, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 111.

[0169] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 104, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 111.

[0170] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 105, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 111.

[0171] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 106, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 111.

[0172] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 107, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 111.

[0173] In some cases, the VH contains an amino acid sequence as shown in SEQ ID NO: 96, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 115.

[0174] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 104, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 115.

[0175] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 105, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 115.

[0176] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 106, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 115.

[0177] In some cases, the VH may contain an amino acid sequence as shown in SEQ ID NO: 107, and the VL may contain an amino acid sequence as shown in SEQ ID NO: 115.

[0178] In this application, the antigen-binding protein may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 167, and the light chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 166.

[0179] In this application, the antigen-binding protein may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 162, and the light chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 159.

[0180] In some cases, the HC contains an amino acid sequence as shown in any one of SEQ ID NO: 124, 132, 133, 134 and 135, and the LC may contain an amino acid sequence as shown in any one of SEQ ID NO: 139 and 143.

[0181] In this application, the antigen-binding protein may comprise HC and LC. HC may comprise VH and a heavy chain constant region. VH may comprise HCDR1-3 and H-FR1-4, where HCDR1 may comprise the amino acid sequence shown in SEQ ID NO.7, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO.153, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO.154. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO.2, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO.12, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.33; VH may comprise the amino acid sequence shown in SEQ ID NO.152; the heavy chain constant region may comprise the amino acid sequence shown in SEQ ID NO.167; and HC may comprise the amino acid sequence shown in SEQ ID NO.162. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.48, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.151. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.159.

[0182] In some cases, the antigen-binding protein may comprise HC and LC. HC may comprise VH and a heavy chain constant region. VH may comprise HCDR1-3 and H-FR1-4, where HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 7, HCDR2 may comprise the amino acid sequence shown in any one of SEQ ID NO. 18, 23, and 24, and HCDR3 may comprise the amino acid sequence shown in any one of SEQ ID NO. 35, 38, and 39. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 2, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 12, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 28, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 43. VH may contain an amino acid sequence as shown in any one of SEQ ID NO. 96, 104, 105, 106, and 107; the heavy chain constant region may contain an amino acid sequence as shown in SEQ ID NO. 167; HC may contain an amino acid sequence as shown in any one of SEQ ID NO. 124, 132, 133, 134, and 135. VL may contain LCDR1-3 and L-FR1-4, LCDR1 may contain an amino acid sequence as shown in SEQ ID NO. 55, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO. 66, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO. 78. L-FR1 may contain amino acid sequences as shown in SEQ ID NO. 48 and 51, L-FR2 may contain an amino acid sequence as shown in SEQ ID NO. 61, L-FR3 may contain an amino acid sequence as shown in SEQ ID NO. 72, and L-FR4 may contain an amino acid sequence as shown in SEQ ID NO. 90. VL may contain the amino acid sequences shown in SEQ ID NO. 111 and 115, the light chain constant region may contain the amino acid sequence shown in SEQ ID NO. 166, and LC may contain the amino acid sequences shown in SEQ ID NO. 139 and 143.

[0183] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000506. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.18, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.35; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.96; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.124. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.111. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. HC may contain the amino acid sequence shown in SEQ ID NO.139.

[0184] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000843. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.23, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.38; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.104; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.132. Furthermore, VL may contain LCDR1-3 and L-FR1-4, LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78; L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90; VL may contain the amino acid sequence shown in SEQ ID NO.111; the light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166; and LC may contain the amino acid sequence shown in SEQ ID NO.139.

[0185] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000844. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.23, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.39; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.105; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.133. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.48, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.111. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.139.

[0186] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000845. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.24, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.38; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.106; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.134. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.48, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.111. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.139.

[0187] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000846. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.24, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.39; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.107; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.135. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.48, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.111. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.139.

[0188] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000847. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.23, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.38; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.104; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.132. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.51, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.115. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.143.

[0189] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000848. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.23, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.39; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.105; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.133. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.51, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.115. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.143.

[0190] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000849. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.24, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.38; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.106; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.134. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.51, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.115. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.143.

[0191] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000850. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.24, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.39; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.107; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.135. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.51, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.115. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.143.

[0192] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000851. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.7, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.18, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.35; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.2, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.12, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.28, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.43; VH may contain the amino acid sequence shown in SEQ ID NO.96; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.124. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.55, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.66, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.78. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.48, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.61, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.72, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.90. VL may contain the amino acid sequence shown in SEQ ID NO.115. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.143.

[0193] In this application, the antigen-binding protein may include at least one HCDR from VH of SEQ ID NO: 156.

[0194] In this application, the antigen-binding protein may comprise HCDR3 from the heavy chain variable region VH of SEQ ID NO: 156, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 34: GGDLLTGPNAFDI (SEQ ID NO. 34). For example, the sequence may be determined according to the definition of Chothia.

[0195] In this application, the antigen-binding protein may comprise HCDR2 from the heavy chain variable region VH of SEQ ID NO: 156, and the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 158: WYX3X4SK (SEQ ID NO. 158); wherein X3 = D or E; X4 = G or A. For example, the sequence may be determined according to the definition of Chothia.

[0196] For example, HCDR2 may contain an amino acid sequence as shown in any of SEQ ID NO: 17, 21 and 22.

[0197] In this application, the antigen-binding protein may comprise HCDR1 from the heavy chain variable region VH of SEQ ID NO: 156, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: GFTFSSY (SEQ ID NO. 6). For example, the sequence may be determined according to the definition of Chothia.

[0198] In this application, the antigen-binding protein may comprise H-FR1, the C-terminus of which is directly or indirectly linked to the N-terminus of HCDR1, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO.1: QVQVVESGGGVVQPGRSLRLSCAAS (SEQ ID NO.1). For example, the sequence may be determined according to the definition of Chothia.

[0199] In this application, the antigen-binding protein may include H-FR2, which is located between HCDR1 and HCDR2, and H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11: DMHWVRQAPGKGLEWVAST (SEQ ID NO.11). For example, the sequence may be determined according to the definition of Chothia.

[0200] In this application, the antigen-binding protein may include H-FR3, which is located between HCDR2 and HCDR3, and H-FR3 may contain the amino acid sequence shown in SEQ ID NO.27: KYYADSVKGRFTISRDNSKNTLYLKMNSLRGDDTAVYYCAK (SEQ ID NO.27). For example, the sequence may be determined according to the definition of Chothia.

[0201] In this application, the antigen-binding protein may comprise H-FR4, the N-terminus of which is directly or indirectly linked to the C-terminus of HCDR3, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.42: WGQGTMVTVSS (SEQ ID NO.42). For example, the sequence may be determined according to the definition of Chothia.

[0202] In this application, the antigen-binding protein may contain VH, and VH may contain the amino acid sequence shown in SEQ ID NO. 156: QVQX4VESGGGVVQPGRSLRLSCAASGFTFSSYDMHWVRQAPGKGLEWVASTWYX5X6SK KYYADSVKGRFTISRDNSKNTLYLX7MNSLRX8X9DTAVYYCAKGGDLLTGPNAFDIWGQG TMVTVSS (SEQ ID NO. 156). Wherein, X4 = L or V, X5 = D or E; X6 = G or A; X7 = K or Q; X8 = G or A; X9 = D or E. For example, the sequence can be determined according to the definition of Chothia.

[0203] For example, VH may contain an amino acid sequence as shown in any one of SEQ ID NO: 95, 99, 100, 101, 102 and 103.

[0204] In this application, the antigen-binding protein may include an antibody heavy chain constant region, and the antibody heavy chain constant region may include a human IgG constant region. In some cases, the human IgG constant region may be derived from the human IgG1 constant region. For example, the antibody heavy chain constant region may include, as in SEQ ID NO.167: ASTKGPSVFPLAPSSKSTSGGTAA

[0205] LGCLVKDYFPEPVTVSW

[0206] NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG The amino acid sequence shown by QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO. 167).

[0207] In this application, the antigen-binding protein may comprise an antibody heavy chain (HC), and the HC comprises an amino acid sequence as shown in SEQ ID NO. 161. For example, the HC may comprise an amino acid sequence as shown in any one of SEQ ID NO: 123, 127, 128, 129, 130, and 131.

[0208] In this application, the antigen-binding protein may comprise at least one LCDR from VL of SEQ ID NO: 155.

[0209] In this application, the antigen-binding protein may comprise LCDR3 from the heavy chain variable region VH of SEQ ID NO: 156, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 157: QQYDX5YX6NT (SEQ ID NO. 157); wherein X5 = M, N, Q, S or T; X6 = E, K, S. For example, the sequence may be determined according to the definition of Chothia.

[0210] In this application, the antigen-binding protein may comprise LCDR2 from the heavy chain variable region VH of SEQ ID NO: 156, and LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 65: KASSLES (SEQ ID NO. 65). For example, the sequence may be determined according to the definition of Chothia.

[0211] In this application, the antigen-binding protein may comprise LCDR1 from the heavy chain variable region VH of SEQ ID NO: 156, and LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 54: RASQSISSWLA (SEQ ID NO. 54). For example, the sequence may be determined according to the definition of Chothia.

[0212] In this application, the antigen-binding protein may include L-FR1, the C-terminus of which is directly or indirectly linked to the N-terminus of LCDR1, and L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47: DIQMTQSPSTLSASVGDRVTITC (SEQ ID NO.47). For example, the sequence may be determined according to the definition of Chothia.

[0213] In this application, the antigen-binding protein may include L-FR2, which is located between LCDR1 and LCDR2, and L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60: WYQQKPGKAPKLLIY (SEQ ID NO.60). For example, the sequence may be determined according to the definition of Chothia.

[0214] In this application, the antigen-binding protein may include L-FR3, which is located between LCDR2 and LCDR3, and L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71: GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC (SEQ ID NO.71). For example, the sequence may be determined according to the definition of Chothia.

[0215] In this application, the antigen-binding protein may comprise L-FR4, the N-terminus of which is directly or indirectly linked to the C-terminus of LCDR3, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO.89: FGQGTKLEIK (SEQ ID NO.89). For example, the sequence may be determined according to the definition of Chothia.

[0216] In this application, the antigen-binding protein may contain a VL, and the VL may contain the amino acid sequence shown in SEQ ID NO. 155: DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLESGVPSRF SGSGSGTEFTLTISSLQPDDFATYYCQQYDX1YX4NTFGQGTKLEIK (SEQ ID NO. 155): where X1 = M, N, Q, S, or T; X4 = E, K, or S. For example, the sequence can be determined according to the definition of Chothia.

[0217] For example, VL may contain an amino acid sequence as shown in any one of SEQ ID NO: 110, 114, 118, 119, 120, 121 and 122.

[0218] In this application, the antigen-binding protein may include an antibody light chain constant region, and the antibody light chain constant region may include a human Igκ constant region. For example, the antibody light chain constant region may include the amino acid sequence shown in SEQ ID NO: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 166).

[0219] In this application, the antigen-binding protein may comprise an antibody light chain (LC), and the LC comprises an amino acid sequence as shown in SEQ ID NO. 160. For example, the LC may comprise an amino acid sequence as shown in any one of SEQ ID NO: 138, 142, 146, 147, 148, 149, and 150.

[0220] In this application, the antigen-binding protein may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO.6, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO.158, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO.34.

[0221] In some cases, antigen-binding proteins may contain HCDR1-3. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in any one of SEQ ID NO.17, 21, and 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34.

[0222] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.6, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.17, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.34.

[0223] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.6, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.21, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.34.

[0224] For example, HCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.6, HCDR2 may contain an amino acid sequence as shown in SEQ ID NO.22, and HCDR3 may contain an amino acid sequence as shown in SEQ ID NO.34.

[0225] In this application, the antigen-binding protein may comprise LCDR1-3. LCDR1 may comprise the amino acid sequence shown in SEQ ID NO.54, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO.157.

[0226] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.77.

[0227] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.81.

[0228] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.84.

[0229] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.85.

[0230] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.86.

[0231] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.87.

[0232] For example, LCDR1 of an antigen-binding protein may contain an amino acid sequence as shown in SEQ ID NO.54, LCDR2 may contain an amino acid sequence as shown in SEQ ID NO.65, and LCDR3 may contain an amino acid sequence as shown in SEQ ID NO.88.

[0233] In this application, the antigen-binding protein may comprise H-FR1-4. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO.164, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO.11, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO.165, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.42.

[0234] For example, H-FR1 may contain the amino acid sequence shown in SEQ ID NO: 1, H-FR2 may contain the amino acid sequence shown in SEQ ID NO: 11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO: 27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO: 42.

[0235] For example, H-FR1 may contain the amino acid sequence shown in SEQ ID NO: 3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO: 11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO: 31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO: 42.

[0236] In this application, the antigen-binding protein may comprise L-FR1-4. L-FR1 may comprise the amino acid sequence shown in SEQ ID NO.47, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO.60, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO.89.

[0237] In this application, the antigen-binding protein may comprise HCDR1-3 and LCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 158, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 34; LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 157.

[0238] In some cases, the HCDR1 of the antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, the HCDR2 may contain the amino acid sequence shown in any one of SEQ ID NO. 17, 21 and 22, and the HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; the LCDR1 may contain the amino acid sequence shown in SEQ ID NO. 54, the LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and the LCDR3 may contain the amino acid sequence shown in any one of SEQ ID NO: 77, 81, 84, 85, 86, 87 and 88.

[0239] For example, HCDR1 of the antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 17, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of the antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 77.

[0240] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 17, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 81.

[0241] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 21, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 77.

[0242] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 77.

[0243] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 81.

[0244] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 84.

[0245] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 85.

[0246] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 86.

[0247] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 87.

[0248] For example, HCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; LCDR1 of an antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO. 88.

[0249] In this application, the antigen-binding protein may comprise H-FR1-4 and L-FR1-4. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 164, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 11, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 165, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 42; L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 47, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 60, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 71, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 89.

[0250] In some cases, H-FR1 may contain the amino acid sequences shown in SEQ ID NO. 1 and 3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO. 11, H-FR3 may contain the amino acid sequences shown in SEQ ID NO. 27 and 31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO. 42; L-FR1 may contain the amino acid sequence shown in SEQ ID NO. 47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO. 60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO. 71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO. 89.

[0251] In this application, the antigen-binding protein may comprise VH and VL. VH may comprise the amino acid sequence shown in SEQ ID NO: 156, and VL may comprise the amino acid sequence shown in SEQ ID NO: 155.

[0252] In some cases, VH may contain an amino acid sequence as shown in any one of SEQ ID NO: 95, 99, 100, 101, 102 and 103, and VL may contain an amino acid sequence as shown in any one of SEQ ID NO: 110, 114, 118, 119, 120, 121 and 122.

[0253] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 95, and VL may contain an amino acid sequence as shown in SEQ ID NO: 110.

[0254] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 99, and VL may contain an amino acid sequence as shown in SEQ ID NO: 110.

[0255] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 100, and VL may contain an amino acid sequence as shown in SEQ ID NO: 110.

[0256] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 101, and VL may contain an amino acid sequence as shown in SEQ ID NO: 110.

[0257] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 110.

[0258] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 103, and VL may contain an amino acid sequence as shown in SEQ ID NO: 110.

[0259] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 95, and VL may contain an amino acid sequence as shown in SEQ ID NO: 114.

[0260] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 100, and VL may contain an amino acid sequence as shown in SEQ ID NO: 114.

[0261] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 114.

[0262] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 103, and VL may contain an amino acid sequence as shown in SEQ ID NO: 114.

[0263] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 118.

[0264] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 119.

[0265] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 120.

[0266] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 121.

[0267] In some cases, VH may contain an amino acid sequence as shown in SEQ ID NO: 102, and VL may contain an amino acid sequence as shown in SEQ ID NO: 122.

[0268] In this application, the antigen-binding protein may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 167, and the light chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 47.

[0269] In this application, the antigen-binding protein may comprise a heavy chain (HC) and a light chain (LC). The HC may comprise the amino acid sequence shown in SEQ ID NO: 161, and the LC may comprise the amino acid sequence shown in SEQ ID NO: 160.

[0270] In some cases, HC may contain an amino acid sequence as shown in any one of SEQ ID NO: 123, 127, 128, 129, 130 and 131, and LC may contain an amino acid sequence as shown in any one of SEQ ID NO: 138, 142, 146, 147, 148, 149 and 150.

[0271] In this application, the antigen-binding protein may contain HC and LC. HC may contain VH and a heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO. 158, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO. 34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO. 164, H-FR2 may contain the amino acid sequence shown in SEQ ID NO. 11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO. 27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO. 42; VH may contain the amino acid sequence shown in SEQ ID NO. 156; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO. 167; and HC may contain the amino acid sequence shown in SEQ ID NO. 161. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.155. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.160.

[0272] In some cases, antigen-binding proteins may contain HC and LC. HC may contain VH and heavy chain constant regions. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in any one of SEQ ID NO.17, 21, and 22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34. H-FR1 may contain any of the amino acid sequences described in SEQ ID NO. 1 and 3; H-FR2 may contain the amino acid sequence shown in SEQ ID NO. 11; H-FR3 may contain the amino acid sequence shown in SEQ ID NO. 27; H-FR4 may contain the amino acid sequence shown in SEQ ID NO. 42; VH may contain any of the amino acid sequences shown in SEQ ID NO: 95, 99, 100, 101, 102, and 103; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO. 167; HC may contain any of the amino acid sequences shown in SEQ ID NO. 123, 127, 128, 129, 130, and 131. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain any one of the amino acid sequences shown in SEQ ID NO.10, 114, 118, 119, 120, 121, and 122. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.47. LC may contain any one of the amino acid sequences shown in SEQ ID NO.138, 142, 146, 147, 148, 149, and 150.

[0273] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000497. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.17, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.1, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.95; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.123. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.110. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.138.

[0274] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000815. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.21, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.1, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.99; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.127. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.110. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.138.

[0275] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000816. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.1, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.100; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.128. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.110. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.138.

[0276] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000817. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.21, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.101; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.129. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.110. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.138.

[0277] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000818. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.110. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.138.

[0278] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000819. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.17, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.103; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.131. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.77. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.110. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.138.

[0279] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000820. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.17, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.1, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.95; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.123. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.81. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.114. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.142.

[0280] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000822. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.1, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.27, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.100; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.128. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.81. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.114. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.142.

[0281] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000824. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.81. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.114. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.142.

[0282] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000825. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.17, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.103; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.131. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.81. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.114. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.142.

[0283] For example, the antigen-binding protein may contain the same HC and LC as antibody PR000832. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.84. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.118. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.146.

[0284] For example, the antigen-binding protein may contain the same HC and LC as antibody PR003833. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.85. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.119. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.146.

[0285] For example, the antigen-binding protein may contain the same HC and LC as antibody PR003834. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.86. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.120. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.148.

[0286] For example, the antigen-binding protein may contain the same HC and LC as antibody PR003835. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.87. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.121. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.149.

[0287] For example, the antigen-binding protein may contain the same HC and LC as antibody PR003836. HC may contain VH and heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4. HCDR1 may contain the amino acid sequence shown in SEQ ID NO.6, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.22, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.34; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.11, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.31, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.42; VH may contain the amino acid sequence shown in SEQ ID NO.102; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.130. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.54, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.65, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.88. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.47, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.71, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.89. VL may contain the amino acid sequence shown in SEQ ID NO.122. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.150.

[0288] In this application, the antigen-binding protein may contain at least one HCDR from VH of SEQ ID NO: 108.

[0289] In this application, the antigen-binding protein may comprise HCDR3 from the heavy chain variable region VH of SEQ ID NO: 108, and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 40: ERSSSFYYYYGMDV (SEQ ID NO. 40). For example, the sequence may be determined according to the definition of Chothia.

[0290] In this application, the antigen-binding protein may comprise HCDR2 from the heavy chain variable region VH of SEQ ID NO: 108, and the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: WYDGSN (SEQ ID NO. 25).

[0291] In this application, the antigen-binding protein may comprise HCDR1 from the heavy chain variable region VH of SEQ ID NO: 108, and the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 10: GFTFYSY (SEQ ID NO. 10). For example, the sequence may be determined according to the definition of Chothia.

[0292] In this application, the antigen-binding protein may comprise H-FR1, the C-terminus of which is directly or indirectly linked to the N-terminus of HCDR1, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO.3: QVQLVESGGGVVQPGRSLRLSCAAS (SEQ ID NO.3). For example, the sequence may be determined according to the definition of Chothia.

[0293] In this application, the antigen-binding protein may include H-FR2, which is located between HCDR1 and HCDR2, and H-FR2 may contain the amino acid sequence shown in SEQ ID NO.15: GMHWVRQTPGKGLEWVALI (SEQ ID NO.15). For example, the sequence may be determined according to the definition of Chothia.

[0294] In this application, the antigen-binding protein may include H-FR3, which is located between HCDR2 and HCDR3, and H-FR3 may contain the amino acid sequence shown in SEQ ID NO.32: NYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAT (SEQ ID NO.32). For example, the sequence may be determined according to the definition of Chothia.

[0295] In this application, the antigen-binding protein may comprise H-FR4, the N-terminus of which is directly or indirectly linked to the C-terminus of HCDR3, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.45: WGQGTTVTVSS (SEQ ID NO.45). For example, the sequence may be determined according to the definition of Chothia.

[0296] In this application, the antigen-binding protein may contain VH, and VH may contain the amino acid sequence shown in SEQ ID NO. 108: QVQLVESGGGVVQPGRSLRLSCAASGFTFYSYGMHWVRQTPGKGLEWVALIWYDGSNNYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCATERSSSFYYYYGMDVWGQGT TVTVSS (SEQ ID NO. 108). For example, the sequence can be determined according to the definition of Chothia.

[0297] In this application, the antigen-binding protein may include an antibody heavy chain constant region, and the antibody heavy chain constant region may include a human IgG constant region. In some cases, the human IgG constant region may be derived from the human IgG1 constant region. For example, the antibody heavy chain constant region may include, as in SEQ ID NO. 167:

[0298] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO.167) amino acid sequence shown.

[0299] In this application, the antigen-binding protein may comprise an antibody heavy chain (HC), which contains an amino acid sequence as shown in SEQ ID NO. 136.

[0300] In this application, the antigen-binding protein may include at least one LCDR from VL of SEQ ID NO: 116.

[0301] In this application, the antigen-binding protein may comprise LCDR3 from the heavy chain variable region VH of SEQ ID NO: 108, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 82: QQLNSYPYT (SEQ ID NO. 82). For example, the sequence may be determined according to the definition of Chothia.

[0302] In this application, the antigen-binding protein may comprise LCDR2 from the heavy chain variable region VH of SEQ ID NO: 108, and LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 69: AASTLQS (SEQ ID NO. 69). For example, the sequence may be determined according to the definition of Chothia.

[0303] In this application, the antigen-binding protein may comprise LCDR1 from the heavy chain variable region VH of SEQ ID NO: 108, and LCDR1 may comprise the amino acid sequence shown in SEQ ID NO: 58: RASQGISSYLA (SEQ ID NO. 58). For example, the sequence may be determined according to the definition of Chothia.

[0304] In this application, the antigen-binding protein may include L-FR1, the C-terminus of which is directly or indirectly linked to the N-terminus of LCDR1, and L-FR1 may contain the amino acid sequence shown in SEQ ID NO. 52: DIQLTQSPSFLSTSVGDRVTITC (SEQ ID NO. 52). For example, the sequence may be determined according to the definition of Chothia.

[0305] In this application, the antigen-binding protein may include L-FR2, which is located between LCDR1 and LCDR2, and L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60: WYQQKPGKAPKLLIY (SEQ ID NO.60). For example, the sequence may be determined according to the definition of Chothia.

[0306] In this application, the antigen-binding protein may include L-FR3, which is located between LCDR2 and LCDR3, and L-FR3 may contain the amino acid sequence shown in SEQ ID NO.75: GVPSRFSGSGSGTEFTLTISSLQPEDFATYYC (SEQ ID NO.75). For example, the sequence may be determined according to the definition of Chothia.

[0307] In this application, the antigen-binding protein may comprise L-FR4, the N-terminus of which is directly or indirectly linked to the C-terminus of LCDR3, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO.93: FGQGTELEIK (SEQ ID NO.93). For example, the sequence may be determined according to the definition of Chothia.

[0308] In this application, the antigen-binding protein may comprise a VL, which may contain an amino acid sequence as shown in SEQ ID NO. 116: DIQLTQSPSFLSTSVGDRVTITCRASQGISSYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQLNSYPYTFGQGTELEIK (SEQ ID NO. 116). For example, the sequence may be determined according to the definition of Chothia.

[0309] In this application, the antigen-binding protein may include an antibody light chain constant region, and the antibody light chain constant region may include a human Igκ constant region. For example, the antibody light chain constant region may include the amino acid sequence shown in SEQ ID NO: 166: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 166).

[0310] In this application, the antigen-binding protein may comprise an antibody light chain (LC), and the LC comprises an amino acid sequence as shown in SEQ ID NO.144.

[0311] In this application, the antigen-binding protein may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO.10, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO.25, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO.40.

[0312] In this application, the antigen-binding protein may comprise LCDR1-3. LCDR1 may comprise the amino acid sequence shown in SEQ ID NO.58, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO.69, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO.82.

[0313] In this application, the antigen-binding protein may comprise H-FR1-4. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO.3, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO.15, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO.32, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO.45.

[0314] In this application, the antigen-binding protein may comprise L-FR1-4. L-FR1 may comprise the amino acid sequence shown in SEQ ID NO.52, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO.60, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO.75, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO.93.

[0315] In this application, the antigen-binding protein may comprise HCDR1-3 and LCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 10, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 25, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 40; LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 58, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 69, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 82.

[0316] In this application, the antigen-binding protein may comprise H-FR1-4 and L-FR1-4. H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 3, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 15, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 32, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 45. L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 52, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 60, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 75, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 93.

[0317] In this application, the antigen-binding protein may comprise VH and VL. VH may comprise the amino acid sequence shown in SEQ ID NO: 108, and VL may comprise the amino acid sequence shown in SEQ ID NO: 116.

[0318] In this application, the antigen-binding protein may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 167, and the light chain constant region may comprise the amino acid sequence shown in SEQ ID NO: 166.

[0319] In this application, the antigen-binding protein may comprise a heavy chain (HC) and a light chain (LC). The HC may comprise the amino acid sequence shown in SEQ ID NO: 136, and the LC may comprise the amino acid sequence shown in SEQ ID NO: 144.

[0320] In this application, the antigen-binding protein may contain HC and LC. HC may contain VH and a heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.10, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.25, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.40; H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.15, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.32, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.45; VH may contain the amino acid sequence shown in SEQ ID NO.108; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.136. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.58, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.69, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.82. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.52, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.75, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.93. VL may contain the amino acid sequence shown in SEQ ID NO.116. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.144.

[0321] For example, the antigen-binding protein may contain the same HC and LC as antibody PR001408. HC may contain VH and a heavy chain constant region. VH may contain HCDR1-3 and H-FR1-4, where HCDR1 may contain the amino acid sequence shown in SEQ ID NO.10, HCDR2 may contain the amino acid sequence shown in SEQ ID NO.25, and HCDR3 may contain the amino acid sequence shown in SEQ ID NO.40. H-FR1 may contain the amino acid sequence shown in SEQ ID NO.3, H-FR2 may contain the amino acid sequence shown in SEQ ID NO.15, H-FR3 may contain the amino acid sequence shown in SEQ ID NO.32, and H-FR4 may contain the amino acid sequence shown in SEQ ID NO.45; VH may contain the amino acid sequence shown in SEQ ID NO.108; the heavy chain constant region may contain the amino acid sequence shown in SEQ ID NO.167; and HC may contain the amino acid sequence shown in SEQ ID NO.136. VL may contain LCDR1-3 and L-FR1-4. LCDR1 may contain the amino acid sequence shown in SEQ ID NO.58, LCDR2 may contain the amino acid sequence shown in SEQ ID NO.69, and LCDR3 may contain the amino acid sequence shown in SEQ ID NO.82. L-FR1 may contain the amino acid sequence shown in SEQ ID NO.52, L-FR2 may contain the amino acid sequence shown in SEQ ID NO.60, L-FR3 may contain the amino acid sequence shown in SEQ ID NO.75, and L-FR4 may contain the amino acid sequence shown in SEQ ID NO.93. VL may contain the amino acid sequence shown in SEQ ID NO.116. The light chain constant region may contain the amino acid sequence shown in SEQ ID NO.166. LC may contain the amino acid sequence shown in SEQ ID NO.144.

[0322] Specific anti-CD73 antibodies

[0323] In this application, the antigen-binding protein can competitively bind to CD73 with a specific anti-CD73 antibody. In some embodiments, the antigen-binding protein of this application may comprise an antibody containing the same amino acid sequence as the specific anti-CD73 antibody.

[0324] In some embodiments, a particular anti-CD73 antibody may comprise at least one HCDR region from the heavy chain variable region VH of SEQ ID NO: 152 and at least one LCDR region from the light chain variable region VL of SEQ ID NO: 151.

[0325] In this application, a specific anti-CD73 antibody may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 7, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 153, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 154. In some cases, a specific anti-CD73 antibody may comprise LCDR1-3. LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 55, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 66, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 78.

[0326] In this application, a specific anti-CD73 antibody may comprise VH and VL. VH may comprise the amino acid sequence shown in SEQ ID NO: 152, and VL may comprise the amino acid sequence shown in SEQ ID NO: 151.

[0327] In this application, a specific anti-CD73 antibody may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise an amino acid sequence as shown in SEQ ID NO: 167, and the light chain constant region may comprise an amino acid sequence as shown in SEQ ID NO: 166.

[0328] In this application, a specific anti-CD73 antibody may comprise a heavy chain (HC) and a light chain (LC). The HC may comprise an amino acid sequence as shown in SEQ ID NO: 162, and the LC may comprise an amino acid sequence as shown in SEQ ID NO: 159.

[0329] In some cases, a particular anti-CD73 antibody may contain at least one HCDR from the heavy chain variable region VH of SEQ ID NO: 156 and at least one LCDR from the light chain variable region VL of SEQ ID NO: 155.

[0330] In this application, a specific anti-CD73 antibody may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 6, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 158, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 34. In some cases, a specific anti-CD73 antibody may comprise LCDR1-3. LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 54, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 65, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 157.

[0331] In this application, a specific anti-CD73 antibody may comprise VH and VL. VH may comprise the amino acid sequence shown in SEQ ID NO: 156, and VL may comprise the amino acid sequence shown in SEQ ID NO: 155.

[0332] In this application, a specific anti-CD73 antibody may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise an amino acid sequence as shown in SEQ ID NO: 167, and the light chain constant region may comprise an amino acid sequence as shown in SEQ ID NO: 166.

[0333] In some cases, a particular anti-CD73 antibody may contain a heavy chain (HC) and a light chain (LC). The HC may contain an amino acid sequence as shown in SEQ ID NO: 161, and the LC may contain an amino acid sequence as shown in SEQ ID NO: 160.

[0334] In some cases, a particular anti-CD73 antibody may comprise at least one HCDR region from the heavy chain variable region VH of SEQ ID NO: 108 and at least one LCDR region from the light chain variable region VL of SEQ ID NO: 116.

[0335] In this application, a specific anti-CD73 antibody may comprise HCDR1-3. HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 10, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 25, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 40. In some embodiments, a specific anti-CD73 antibody may comprise LCDR1-3. LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 58, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 69, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 82.

[0336] In this application, a specific anti-CD73 antibody may contain VH and VL. VH may contain the amino acid sequence shown in SEQ ID NO: 108, and VL may contain the amino acid sequence shown in SEQ ID NO: 116.

[0337] In this application, a specific anti-CD73 antibody may comprise a heavy chain constant region and a light chain constant region. The heavy chain constant region may comprise an amino acid sequence as shown in SEQ ID NO: 167, and the light chain constant region may comprise an amino acid sequence as shown in SEQ ID NO: 166.

[0338] In this application, a specific anti-CD73 antibody may comprise a heavy chain (HC) and a light chain (LC). The HC may comprise an amino acid sequence as shown in SEQ ID NO: 136, and the LC constant region may comprise an amino acid sequence as shown in SEQ ID NO: 144.

[0339] The antigen-binding protein disclosed herein may also include homologs or variants thereof having substantially the same function / properties. In some embodiments, the homolog or variant may be a polypeptide that differs from the antigen-binding protein by at least one amino acid. For example, the homolog or variant may be a polypeptide obtained by adding, deleting, or substituting one or more amino acids (e.g., 1-50, 1-40, 1-30, 1-20, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) to the antigen-binding protein.

[0340] In some cases, antigen-binding proteins can be further optimized through germlineation and PTM removal. The term "germlineation" refers to the process of reversing a supersomal mutation of an antibody into a corresponding germline sequence. The term "PTM removal" refers to the process of mutating the PTM motif to reduce the risk of sequence instability.

[0341] Post-translational modifications (PTMs) are widely observed in proteins expressed in mammalian cells, potentially leading to molecular instability and heterogeneity. To reduce sequence instability, PTM motifs can be removed through mutation. PTM removal sites can be isomerized motifs, such as DG in the CDR. In some cases, PTM removal sites may include one or more sites selected from D54, G55, D99, and / or G100 in the HCDR. In some cases, PTM removal in D54 may be D54E. In some cases, PTM removal in G55 may be G55A. In some cases, PTM removal in D99 may be D99E. In some cases, PTM removal in G100 may be G100A.

[0342] PTM-removed antibodies can possess similar or higher chemical stability or biological activity. In some cases, PTM-removed antibodies may exhibit similar or higher binding affinity, enzymatic activity, T-cell activation activity, tumor suppressor activity, and / or serum stability. For example, PTM-removed antibodies (e.g., PR000844, PR000846) have comparable binding affinity to PTM antibodies (e.g., PR000506).

[0343] In some cases, a homolog or variant can be a polypeptide that has at least 80% sequence identity with an antigen-binding protein. For example, a homolog or variant can be a polypeptide that has at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or higher) sequence identity with an antibody or antigen-binding protein.

[0344] As used in the context of describing the polypeptide sequences identified herein, the term "sequence identity percentage (%)" generally refers to the percentage of amino acid residues or nucleotides in the query sequence that are identical to residues in a second reference polypeptide sequence or a portion thereof after sequence alignment, with vacancies (GAPS) introduced where necessary to achieve the maximum sequence identity percentage, and without considering any conserved substitutions as part of the sequence identity. Alignments used to determine the amino acid / nucleotide sequence identity percentage can be performed in various ways known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, NEEDLE, or Megalign (DNASTAR). Those skilled in the art can determine suitable parameters for measuring the alignment, including any algorithm required to achieve maximum alignment across the full length of the sequences being compared. The identity percentage can be determined over the entire length of the identified polypeptide / polynucleotide sequence or over a shorter length, such as the length of a fragment obtained from a larger identified polypeptide / polynucleotide sequence. It should be understood that any fragment length supported by the sequences shown herein in tables, figures, or sequence listings can be used as a length for which a measurable identity percentage can be determined.

[0345] Nucleic acids, vectors, and cells

[0346] On the other hand, this application provides one or more isolated nucleic acid molecules encoding isolated antigen-binding proteins.

[0347] The isolated nucleic acid may contain one or more nucleic acid molecules, each encoding an antigen-binding protein. For example, the isolated nucleic acid may contain at least two nucleic acid molecules, one encoding an antibody heavy chain or a fragment thereof, and the other encoding an antibody light chain or a fragment thereof.

[0348] One or more isolated nucleic acids can be synthesized using recombinant techniques well known in the art. For example, one or more isolated nucleic acids can be synthesized using an automated DNA synthesizer. Standard recombinant DNA and molecular cloning techniques are documented in Sambrook, J., Fritsch, EF, and Maniatis, T. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, (1989) (Maniatis) and TJ Silhavy, MLBennan, and LW Enquist, Experiments with Gene Fusions, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1984) and Ausubel, FM, et al., Current Protocols in Molecular Biology, pub. by Greene Publishing Assoc. and Wiley-Interscience (1987). In short, target nucleic acids can be prepared from genomic DNA fragments, cDNA, and RNA, all of which can be extracted directly from cells or recombinantly generated through various amplification methods, including but not limited to PCR and RT-PCR.

[0349] Direct chemical synthesis of nucleic acids typically involves the sequential addition of 3'-blocked and 5'-blocked nucleotide monomers to the terminal 5'-hydroxyl group of a growing nucleotide polymer chain, wherein each addition is achieved by nucleophilic attack on the terminal 5'-hydroxyl group of the added monomer (typically a phosphorus derivative, such as a triphosphate, phosphoramide, etc.) at the 3'-position. See, for example, Matteuci et al., Tet. Lett. 521:719 (1980); U.S. Patent 4,500,707, Caruthers et al.; and U.S. Patents 5,436,327 and 5,700,637, Southern et al.

[0350] On the other hand, this disclosure provides one or more vectors that contain one or more isolated nucleic acid molecules.

[0351] The vector can be any linear nucleic acid, plasmid, phage particle, granule, RNA vector, viral vector, etc. Non-limiting examples of viral vectors may include retroviruses, adenoviruses, and adeno-associated viruses. In some embodiments, the vector is an expression vector, such as a plasmid.

[0352] Expression vectors may be suitable for specific types of host cells but not for others. For example, an expression vector can be introduced into a host organism, and then the viability of the host organism and the expression of any genes / polynucleotides contained in the vector can be monitored.

[0353] Expression vectors may also contain one or more selectable marker genes that, upon expression, confer one or more phenotypic traits, which can be used to select or otherwise identify host cells carrying the expression vector. Non-restrictive examples of suitable selectable markers for eukaryotic cells include dihydrofolate reductase and neomycin resistance.

[0354] Target vectors can be stably or transiently transfected into host cells using a variety of established techniques. For example, one method involves calcium chloride treatment, where the expression vector is introduced via a calcium precipitate. Other salts, such as calcium phosphate, can also be used following similar steps. Additionally, electroporation (i.e., applying an electric current to increase cell permeability to nucleic acids) can be used. Other examples of transformation methods include microinjection, DEAE-mediated transformation, and heat shock in the presence of lithium acetate. Lipid complexes, liposomes, and dendritic polymers can also be used to transfect host cells.

[0355] On the other hand, this application provides a cell (e.g., an isolated cell, such as a host cell) containing one or more isolated nucleic acid molecules of this application or one or more vectors of this application. The cell can express the antigen-binding protein of this application. The cell can be a eukaryotic cell or a prokaryotic cell. Suitable cells can be transformed or transfected with the nucleic acid or vector of this application and used to express and / or secrete the antigen-binding protein. For example, the cell can be HEK293 cells, other bacterial host cells, yeast cells, or various higher-order eukaryotic cells.

[0356] On the other hand, this disclosure provides a method for generating the antigen-binding protein of this disclosure, comprising culturing the cells of this disclosure under conditions capable of expressing the antigen-binding protein. The method may optionally further include harvesting the antigen-binding protein of this disclosure.

[0357] Pharmaceutical compositions, uses and methods

[0358] In another aspect, this disclosure provides pharmaceutical compositions comprising isolated antigen-binding proteins, one or more nucleic acid molecules, one or more carriers, and / or cells, as well as pharmaceutically acceptable carriers.

[0359] As used herein, "pharmaceutically acceptable carriers" include any and all preservatives, humectants, emulsifiers, and dispersants. The presence of microorganisms can be prevented by sterilization procedures and by the addition of various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenolic sorbic acid, etc. Isotonic agents, such as sugars, sodium chloride, etc., may also be added to the composition. Furthermore, the absorption of injectable drug forms can be prolonged by adding agents that delay absorption, such as aluminum monostearate and gelatin.

[0360] Pharmaceutical compositions must generally be sterile and stable under manufacturing and storage conditions. Compositions can be formulated as solutions, microemulsions, liposomes, or other ordered structures suitable for high drug concentrations. The carrier can be a solvent or dispersion medium comprising, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. For example, appropriate flowability can be maintained by using coatings such as lecithin, maintaining the desired particle size in the case of dispersions, and by using surfactants. In many cases, isotonic agents, such as sugars, polyols like mannitol, sorbitol, or sodium chloride, are preferably included in the composition. Extended absorption of injectable compositions can be achieved by adding agents that delay absorption, such as monostearate and gelatin.

[0361] The pharmaceutical compositions disclosed herein may include other compounds, drugs, and / or agents for the treatment of cancer. Such compounds, drugs, and / or agents may include, for example, chemotherapeutic agents, small molecule drugs, or antibodies that stimulate an immune response against a given cancer. In some cases, the pharmaceutical composition may include one or more agents, such as those listed in the combination therapy section. For example, a combination therapy may include the anti-CD73 antibody described herein combined with at least one other anticancer and / or T-cell stimulating (e.g., activating) agent. Examples of therapeutic agents that may be used in combination therapies are described in more detail below in the section concerning the use of the antibodies described herein.

[0362] Isolated antigen-binding proteins and / or pharmaceutical compositions can be used to prevent, alleviate, and / or treat tumors. The tumors are solid tumors and / or hematologic malignancies.

[0363] On the other hand, this disclosure further provides a method for treating tumors, comprising preventing, alleviating, and / or treating tumors, the method comprising administering an isolated antigen-binding protein to a subject in need of such treatment, thereby inhibiting or reducing tumor growth and / or causing tumor regression. The isolated antigen-binding protein may be used alone to inhibit the growth of cancerous tumors. Alternatively, the isolated antigen-binding protein may be used in combination with another agent known in the art, such as other immunogenic agents, standard cancer treatments, or other antibodies. Therefore, this application provides a method for preventing, alleviating, and / or treating tumors, for example, by inhibiting the growth of tumor cells in a subject, comprising administering to a subject a therapeutically effective amount of the antigen-binding protein described herein or its antigen-binding portion.

[0364] This disclosure also discloses the following implementation plan:

[0365] 1. The isolated antigen-binding protein described above has one or more of the following properties:

[0366] a. It can bind human CD73 and cynomolgus monkey CD73 with considerable affinity;

[0367] b. It can inhibit the 5′ exonuclease activity of CD73;

[0368] c. It can mediate the internalization of CD73;

[0369] d. It can relieve the inhibitory effect of adenosine on T cell proliferation;

[0370] e. Maintain a relatively stable serum concentration for at least 15 days; and

[0371] f. It can inhibit tumor growth and / or tumor cell proliferation.

[0372] 2. The isolated antigen-binding protein of embodiment 1, comprising at least one HCDR region from the heavy chain variable region VH of SEQ ID NO: 152.

[0373] 3. An isolated antigen-binding protein of any one of embodiments 1-2, comprising HCDR3 from the heavy chain variable region VH of SEQ ID NO: 152.

[0374] 4. An isolated antigen-binding protein of any one of embodiments 1-3, comprising HCDR2 from the heavy chain variable region VH of SEQ ID NO: 152.

[0375] 5. An isolated antigen-binding protein of any one of embodiments 1-4, comprising HCDR1 from the heavy chain variable region VH of SEQ ID NO: 152.

[0376] 6. The isolated antigen-binding protein of any one of embodiments 1-5, wherein the VH comprises the amino acid sequence shown in any one of SEQ ID NO: 96, 104, 105, 106 and 107.

[0377] 7. An isolated antigen-binding protein of any one of embodiments 1-6, comprising at least one LCDR region from the light chain variable region VL of SEQ ID NO: 151.

[0378] 8. An isolated antigen-binding protein of any one of embodiments 1-7, comprising LCDR3 from the light chain variable region VL of SEQ ID NO: 151.

[0379] 9. An isolated antigen-binding protein of any one of embodiments 1-8, comprising LCDR2 from the light chain variable region VL of SEQ ID NO: 151.

[0380] 10. An isolated antigen-binding protein of any one of embodiments 1-9, comprising LCDR1 from the light chain variable region VL of SEQ ID NO: 151.

[0381] 11. An isolated antigen-binding protein of any one of embodiments 1-10, wherein the VL comprises the amino acid sequence shown in any one of SEQ ID NO: 111 and 115.

[0382] 12. An isolated antigen-binding protein according to any one of embodiments 1-11, comprising an antibody or an antigen-binding fragment thereof.

[0383] 13. The isolated antigen-binding protein of embodiment 12, wherein the antigen-binding fragment comprises Fab, Fab′, F(ab)2, Fv fragment, F(ab′)2, scFv, di-scFv and / or dAb.

[0384] 14. An isolated antigen-binding protein of any one of embodiments 1-13, wherein the antibody is selected from monoclonal antibodies, chimeric antibodies, humanized antibodies and fully human antibodies.

[0385] 15. An isolated antigen-binding protein of any one of embodiments 3-14, wherein the HCDR3 comprises the amino acid sequence shown in any one of SEQ ID NO: 35, 38 and 39.

[0386] 16. An isolated antigen-binding protein of any one of embodiments 4-15, wherein the HCDR2 comprises the amino acid sequence shown in any one of SEQ ID NO: 18, 23 and 24.

[0387] 17. An isolated antigen-binding protein according to any one of embodiments 1-16, comprising a light chain variable region VL, wherein the VL of the antigen-binding protein comprises frame regions L-FR1, L-FR2, L-FR3 and L-FR4.

[0388] 18. The isolated antigen-binding protein of embodiment 17, wherein the C-terminus of said L-FR1 is directly or indirectly linked to the N-terminus of said LCDR1, and said L-FR1 comprises an amino acid sequence as shown in SEQ ID NO: 163.

[0389] 19. An isolated antigen-binding protein of any one of embodiments 17-18, wherein the L-FR1 comprises an amino acid sequence as shown in any one of SEQ ID NO: 48 and 51.

[0390] 20. An isolated antigen-binding protein of any one of embodiments 17-19, wherein the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises an amino acid sequence as shown in SEQ ID NO: 61.

[0391] 21. An isolated antigen-binding protein of any one of embodiments 17-20, wherein the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 comprises an amino acid sequence as shown in SEQ ID NO: 72.

[0392] 22. An isolated antigen-binding protein of any one of embodiments 17-21, wherein the N-terminus of said L-FR4 is directly or indirectly linked to the C-terminus of said LCDR3, and said L-FR4 comprises an amino acid sequence as shown in SEQ ID NO: 90.

[0393] 23. An isolated antigen-binding protein of any one of embodiments 1-22, wherein the VL of the antigen-binding protein comprises an amino acid sequence as shown in SEQ ID NO: 151.

[0394] 24. An isolated antigen-binding protein of any one of embodiments 1-23, wherein the VL of the antigen-binding protein comprises an amino acid sequence as shown in any one of SEQ ID NO: 111 and 115.

[0395] 25. An isolated antigen-binding protein according to any one of embodiments 1-24, comprising an antibody light chain constant region, wherein the antibody light chain constant region comprises a human Igκ constant region.

[0396] 26. The isolated antigen-binding protein of embodiment 25, wherein the constant region of the antibody light chain comprises an amino acid sequence as shown in SEQ ID NO: 166.

[0397] 27. An isolated antigen-binding protein of any one of embodiments 1-26, comprising an antibody light chain LC, wherein the LC comprises an amino acid sequence as shown in SEQ ID NO: 159.

[0398] 28. An isolated antigen-binding protein of any one of embodiments 1-27, comprising an antibody light chain LC, wherein the LC comprises an amino acid sequence shown in any one of SEQ ID NO: 139 and 143.

[0399] 29. An isolated antigen-binding protein according to any one of embodiments 1-28, comprising a heavy chain variable region VH, wherein the VH of the antigen-binding protein comprises frame regions H-FR1, H-FR2, H-FR3 and H-FR4.

[0400] 30. The isolated antigen-binding protein of embodiment 29, wherein the C-terminus of said H-FR1 is directly or indirectly linked to the N-terminus of said HCDR1, and said H-FR1 comprises the amino acid sequence shown in SEQ ID NO: 2.

[0401] 31. An isolated antigen-binding protein of any one of embodiments 29-30, wherein the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 comprises an amino acid sequence as shown in SEQ ID NO: 12.

[0402] 32. An isolated antigen-binding protein of any one of embodiments 29-31, wherein the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises an amino acid sequence as shown in SEQ ID NO: 28.

[0403] 33. An isolated antigen-binding protein of any one of embodiments 29-32, wherein the N-terminus of said H-FR4 is directly or indirectly linked to the C-terminus of said HCDR3, and said H-FR4 comprises the amino acid sequence shown in SEQ ID NO: 43.

[0404] 34. An isolated antigen-binding protein of any one of embodiments 1-33, wherein the VH of the antigen-binding protein comprises an amino acid sequence as shown in SEQ ID NO: 152.

[0405] 35. An isolated antigen-binding protein of any one of embodiments 1-34, wherein the VH of the antigen-binding protein comprises an amino acid sequence as shown in any one of SEQ ID NO: 96, 104, 105, 106 and 107.

[0406] 36. An isolated antigen-binding protein according to any one of embodiments 1-35, comprising an antibody heavy chain constant region, wherein the antibody heavy chain constant region comprises a human IgG constant region.

[0407] 37. An isolated antigen-binding protein according to any one of embodiments 1-36, comprising an antibody heavy chain constant region, wherein the antibody heavy chain constant region comprises a human IgG1 constant region.

[0408] 38. The isolated antigen-binding protein of embodiment 37, wherein the constant region of the antibody heavy chain contains an amino acid sequence as shown in SEQ ID NO: 167.

[0409] 39. An isolated antigen-binding protein of any one of embodiments 1-38, comprising an antibody heavy chain HC, wherein said HC comprises an amino acid sequence as shown in SEQ ID NO: 162.

[0410] 40. An isolated antigen-binding protein of any one of embodiments 1-39, comprising an antibody heavy chain HC, wherein the HC comprises an amino acid sequence as shown in any one of SEQ ID NO: 124, 132, 133, 134 and 135.

[0411] 41. One or more isolated nucleic acid molecules encoding the isolated antigen-binding protein of any one of embodiments 1-40.

[0412] 42. One or more vectors comprising the isolated one or more isolated nucleic acid molecules of embodiment 41.

[0413] 43. A cell comprising the nucleic acid molecule or molecule of the isolated embodiment 41, or one or more carriers of embodiment 42.

[0414] 44. A method for producing an isolated antigen-binding protein of any one of embodiments 1-40, comprising culturing cells of embodiment 43 under conditions capable of expressing the isolated antigen-binding protein of any one of embodiments 1-40.

[0415] 45. A pharmaceutical composition comprising an isolated antigen-binding protein of any one of embodiments 1-40, one or more nucleic acid molecules of embodiment 41, one or more carriers of embodiment 42 and / or cells of embodiment 43, and optionally, a pharmaceutically acceptable adjuvant.

[0416] 46. ​​The isolated antigen-binding protein of any one of embodiments 1-40, the isolated one or more nucleic acid molecules of embodiment 41, the one or more carriers of embodiment 42, the cells of embodiment 43, and / or the pharmaceutical composition of embodiment 45 for the production of a medicament for the prevention, mitigation, and / or treatment of tumors.

[0417] 47. Use according to embodiment 46, wherein the tumor includes solid tumors and / or hematologic malignancies.

[0418] 48. A method for preventing, alleviating and / or treating tumors, comprising administering to a subject in need an isolated antigen-binding protein of any one of embodiments 1-40.

[0419] This disclosure also discloses the following implementation plan:

[0420] 1. An isolated antigen-binding protein comprising any one of the HCDR1, HCDR2 and / or HCDR3 regions of the variable region VH of the antibody heavy chain shown in SEQ ID NO:168.

[0421] 2. The isolated antigen-binding protein according to embodiment 1, further comprising any one of the LCDR1, LCDR2 and / or LCDR3 regions of the antibody light chain variable region shown in SEQ ID NO: 169.

[0422] 3. The isolated antigen-binding protein according to any one of embodiments 1-2, having one or more of the following properties:

[0423] a. It can bind human CD73 and cynomolgus monkey CD73 with considerable affinity;

[0424] b. It can inhibit the 5′ exonuclease activity of CD73;

[0425] c. It can regulate the internalization of CD73;

[0426] d. Can promote T cell proliferation;

[0427] e. Maintain a relatively stable serum concentration for at least 15 days; and

[0428] f. It can inhibit tumor growth and / or tumor cell proliferation.

[0429] 4. An isolated antigen-binding protein according to any one of embodiments 1-3, comprising an antibody or an antigen-binding fragment thereof.

[0430] 5. The isolated antigen-binding protein according to embodiment 4, wherein the antigen-binding fragments include Fab, Fab', F(ab)2, Fv fragments, F(ab')2, scFv, di-scFv and / or dAb.

[0431] 6. The isolated antigen-binding protein according to any one of embodiments 4-5, wherein the antibody is selected from monoclonal antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.

[0432] 7. The isolated antigen-binding protein according to any one of embodiments 1-6, comprising any one of the HCDR1, HCDR2 and / or HCDR3 regions of the heavy chain variable region VH shown in SEQ ID NO: 152.

[0433] 8. The isolated antigen-binding protein according to embodiment 7, wherein the VH comprises the amino acid sequence shown in any one of SEQ ID NO: 96, 104, 105, 106 and 107.

[0434] 9. The isolated antigen-binding protein according to any one of embodiments 7-8, comprising any one of the LCDR1, LCDR2 and / or LCDR3 regions of the light chain variable region VL shown in SEQ ID NO: 151.

[0435] 10. The isolated antigen-binding protein according to embodiment 9, wherein the VL comprises any one of the amino acid sequences shown in SEQ ID NO: 111 and 115.

[0436] 11. The isolated antigen-binding protein according to any one of embodiments 7-10, wherein the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:154.

[0437] 12. The isolated antigen-binding protein according to any one of embodiments 7-11, wherein the HCDR3 comprises the amino acid sequence shown in any one of SEQ ID NO: 35, 38 and 39.

[0438] 13. The isolated antigen-binding protein according to any one of embodiments 7-12, wherein the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:153.

[0439] 14. The isolated antigen-binding protein according to any one of embodiments 7-13, wherein the HCDR2 comprises the amino acid sequence shown in any one of SEQ ID NO: 18, 23 and 24.

[0440] 15. The isolated antigen-binding protein according to any one of embodiments 7-14, wherein the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:7.

[0441] 16. The isolated antigen-binding protein according to any one of embodiments 7-15, comprising a heavy chain variable region VH, wherein the VH comprises the amino acid sequence shown in SEQ ID NO:152.

[0442] 17. The isolated antigen-binding protein according to embodiment 16, wherein the VH comprises the amino acid sequence shown in any one of SEQ ID 96, 104, 105, 106 and 107.

[0443] 18. The isolated antigen-binding protein according to any one of embodiments 7-17, comprising an antibody heavy chain HC, said HC comprising an amino acid sequence shown in any one of SEQ ID NO: 124, 132, 133, 134, and 135.

[0444] 19. The isolated antigen-binding protein according to any one of embodiments 9-18, wherein the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:78.

[0445] 20. The isolated antigen-binding protein according to any one of embodiments 9-19, wherein the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:66.

[0446] 21. The isolated antigen-binding protein according to any one of embodiments 9-20, wherein the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:55.

[0447] 22. The isolated antigen-binding protein according to any one of embodiments 9-21, comprising a light chain variable region VL, said VL comprising the amino acid sequence shown in SEQ ID NO:151.

[0448] 23. The isolated antigen-binding protein according to any one of embodiments 9-22, wherein the VL comprises the amino acid sequence shown in any one of SEQ ID NO: 111 and 115.

[0449] 24. The isolated antigen-binding protein according to any one of embodiments 7-23, comprising an antibody light chain LC, said LC comprising an amino acid sequence shown in any one of SEQ ID NO: 139 and 143.

[0450] 25. The isolated antigen-binding protein according to any one of embodiments 1-6, comprising any one of the HCDR1, HCDR2 and / or HCDR3 regions of the heavy chain variable region VH shown in SEQ ID NO: 156.

[0451] 26. The isolated antigen-binding protein according to embodiment 25, wherein the VH comprises an amino acid sequence shown in any one of SEQ ID NO: 95, 99, 100, 101, 102, and 103.

[0452] 27. The isolated antigen-binding protein according to any one of embodiments 25-26, comprising any one of the LCDR1, LCDR2 and / or LCDR3 regions of the light chain variable region VL shown in SEQ ID NO: 155.

[0453] 28. The isolated antigen-binding protein according to embodiment 27, wherein the VL comprises the amino acid sequence shown in any one of SEQ ID NO: 110, 114, 118, 119, 120, 121, and 122.

[0454] 29. The isolated antigen-binding protein according to any one of embodiments 25-28, wherein the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:34.

[0455] 30. The isolated antigen-binding protein according to any one of embodiments 25-29, wherein the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:158.

[0456] 31. The isolated antigen-binding protein according to any one of embodiments 25-30, wherein the HCDR2 is selected from the amino acid sequence comprising any one of SEQ ID NO: 17, 21 and 22.

[0457] 32. The isolated antigen-binding protein according to any one of embodiments 25-31, wherein the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:6.

[0458] 33. The isolated antigen-binding protein according to any one of embodiments 25-32, comprising a heavy chain variable region VH, wherein the VH comprises the amino acid sequence shown in SEQ ID NO:156.

[0459] 34. The isolated antigen-binding protein according to embodiment 33, wherein the VH comprises an amino acid sequence shown in any one of SEQ ID NO: 95, 99, 100, 101, 102 and 103.

[0460] 35. The isolated antigen-binding protein according to any one of embodiments 25-34, comprising an antibody heavy chain HC, wherein the HC comprises an amino acid sequence represented by any one of SEQ ID NO: 123, 127, 128, 129, 130 and 131.

[0461] 36. The isolated antigen-binding protein according to any one of embodiments 25-35, wherein the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:157.

[0462] 37. The isolated antigen-binding protein according to any one of embodiments 25-36, wherein the LCDR3 comprises the amino acid sequence shown in any one of SEQ ID NO: 77, 81, 84, 85, 86, 87 and 88.

[0463] 38. The isolated antigen-binding protein according to any one of embodiments 27-37, wherein the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:65.

[0464] 39. The isolated antigen-binding protein according to any one of embodiments 27-38, wherein the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:54.

[0465] 40. The isolated antigen-binding protein according to any one of embodiments 27-39, comprising a light chain variable region VL, wherein the VL comprises the amino acid sequence shown in SEQ ID NO:155.

[0466] 41. The isolated antigen-binding protein according to embodiment 40, wherein the VL comprises the amino acid sequence shown in any one of SEQ ID Nos: 110, 114, 118, 119, 120, 121 and 122.

[0467] 42. The isolated antigen-binding protein according to any one of embodiments 25-41, comprising an antibody light chain LC, wherein the LC comprises an amino acid sequence shown in any one of SEQ ID NO: 138, 142, 146, 147, 148, 149 and 150.

[0468] 43. The isolated antigen-binding protein according to any one of embodiments 1-6, comprising any one of the HCDR1, HCDR2 and / or HCDR3 regions from the heavy chain variable region VH shown in SEQ ID NO:108.

[0469] 44. The isolated antigen-binding protein according to embodiment 43 comprises any one of the LCDR1, LCDR2 and / or LCDR3 regions from the light chain variable region VL shown in SEQ ID NO:116.

[0470] 45. The isolated antigen-binding protein according to any one of embodiments 43-44, wherein the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:40.

[0471] 46. ​​The isolated antigen-binding protein according to any one of embodiments 43-45, wherein the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:25.

[0472] 47. The isolated antigen-binding protein according to any one of embodiments 43-46, wherein the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:10.

[0473] 48. The isolated antigen-binding protein according to any one of embodiments 43-47, comprising a heavy chain variable region VH, wherein the VH comprises the amino acid sequence shown in SEQ ID NO: 108.

[0474] 49. The isolated antigen-binding protein according to any one of embodiments 43-48, comprising an antibody heavy chain HC, wherein the HC comprises the amino acid sequence shown in SEQ ID NO:136.

[0475] 50. The isolated antigen-binding protein according to any one of embodiments 44-49, wherein the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:82.

[0476] 51. The isolated antigen-binding protein according to any one of embodiments 44-50, wherein the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:69.

[0477] 52. The isolated antigen-binding protein according to any one of embodiments 44-51, wherein the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:58.

[0478] 53. The isolated antigen-binding protein according to any one of embodiments 44-52, comprising a light chain variable region VL, said VL comprising the amino acid sequence shown in SEQ ID NO:116.

[0479] 54. The isolated antigen-binding protein according to any one of embodiments 43-53, comprising an antibody light chain LC, said LC comprising the amino acid sequence shown in SEQ ID NO:144.

[0480] 55. One or more isolated nucleic acid molecules encoding the isolated antigen-binding protein described in any one of embodiments 1-54.

[0481] 56. One or more vectors comprising one or more isolated nucleic acid molecules as described in embodiment 55.

[0482] 57. A cell comprising one or more isolated nucleic acid molecules as described in embodiment 55 or one or more vectors as described in embodiment 56.

[0483] 58. A method for preparing the isolated antigen-binding protein according to any one of embodiments 1-54, the method comprising culturing the cells described in embodiment 57 under conditions that cause the isolated antigen-binding protein according to any one of embodiments 1-54 to be expressed.

[0484] 59. A pharmaceutical composition comprising the isolated antigen-binding protein of any one of embodiments 1-54, one or more isolated nucleic acid molecules of embodiment 55, the carrier of embodiment 56, and / or the cell of embodiment 57, and optionally a pharmaceutically acceptable carrier.

[0485] 60. The use of the isolated antigen-binding protein of any one of embodiments 1-54, one or more nucleic acid molecules of embodiment 55, one or more carriers of embodiment 56, the cell of embodiment 57, and / or the pharmaceutical composition of embodiment 59 in the preparation of a medicament for the prevention, relief, and / or treatment of tumors.

[0486] 61. The use described in embodiment 60, wherein the tumor includes solid tumors and / or hematologic malignancies.

[0487] 62. A method for preventing, alleviating and / or treating tumors, the method comprising administering to a subject in need the isolated antigen-binding protein as described in any one of embodiments 1-54.

[0488] 63. A method for inhibiting the 5′ exonuclease activity of CD73, the method comprising administering the isolated antigen-binding protein as described in any one of embodiments 1-54.

[0489] 64. A method for regulating CD73 internalization, the method comprising administering the isolated antigen-binding protein as described in any one of embodiments 1-54. Example

[0490] The following embodiments are provided to provide a complete disclosure and illustration to those skilled in the art regarding how to implement and use the invention, and are not intended to limit the scope as the inventors believe, nor are they intended to represent all or only the experiments of the invention. Every effort has been made to ensure the accuracy of the figures used (e.g., quantities, temperatures, etc.), but some experimental errors and biases should still be taken into account. Unless otherwise stated, parts are parts by weight, molecular weight is weight-average molecular weight, temperature is degrees Celsius, and pressure is atmospheric pressure or near atmospheric pressure. Standard abbreviations may be used, such as bp, base pairs; kb, kilobases; pl, picoliter; s or sec; minutes; h or hr, hours; aa, amino acids; nt, nucleotides; im, intramuscular (ly); ip, intraperitoneal (ly); sc, subcutaneous injection, etc.

[0491] Example 1: Production of anti-CD73 antibody

[0492] immunity

[0493] In immunization using Harbor H2L2 mice, CD73 protein was used as an immunogen to generate anti-CD73 antibodies. Each mouse was given a first booster of 50 μg protein, along with an adjuvant (Sigma, S6322), via subcutaneous and intraperitoneal injections. Subsequent boosters were 25 μg protein. This immunization was performed every two weeks for a total of six times. Final immunization was performed via intraperitoneal injection of the immunogen diluted with PBS. Serum titers against human CD73 were tested using ELISA and FACS. Mouse serum was sampled and titrated at specified time points for ELISA and FACS analysis to test binding to CD73 or CD73-stable cell lines. Good serum titers were observed in mice in the protein-immunized group.

[0494] Hybridoma

[0495] Using a cell fusion generator (BEX-LF301), mouse spleen cells isolated from immunized mice were fused with the mouse myeloma cell line SP2 / 0 (ATCC, CRL-1581) via an electric field-based electroporator to obtain hybridomas. Typically, human anti-CD73 antibodies were screened in each well 9-14 days post-fusion. After hybridoma subcloning, monoclonal 38H6, exhibiting good binding activity to CD73, was selected for sequencing. The heavy chain variable region and light chain variable region were identified and obtained. The heavy chain variable region and light chain variable region were then synthesized and cloned into plasmids encoding the human IgG1 constant region and the human Igκ region, respectively.

[0496] Example 2 Antibody Production and Purification

[0497] The recombinant plasmid encoding the target antibody was transiently co-transfected into HEK293-6E or 293-F cell cultures using PEI (Polyscience, 24885). 30 μg of plasmid was mixed with 120 μL of PEI and incubated at room temperature for 15 minutes. Next, the mixture was added dropwise to a 1×10⁻⁶ ppm solution. 6 Cells were suspended in Opti-MEM 293 cells at a concentration of 1 / mL. After transfection, the cells were cultured at 37°C and 5% CO2 using a shaker at 120 rpm. The cell culture supernatant collected on days 6-7 was used for purification.

[0498] Six to seven days post-transfection, the supernatant containing the target antibody was harvested by centrifugation and filtration. The monoclonal antibody was purified using a pre-packed rProtein A (GE, 17-1279-02) column (Bio-Rad, 7311550). Protein A columns were prepared by packing 0.2 mL of rProtein A resin into a column and washing with 10 column volumes of ddH2O and 10 column volumes of PBS. The cell culture supernatant was passed through the column and washed with 10 column volumes of PBS. Proteins were then eluted with 8 column volumes of elution buffer (Thermo, 21004) and immediately mixed with 640 μL of neutralization buffer (1 M Tris-HCl, pH 9.0; Teknova, T1090). The buffer solution was centrifuged at 3800 rpm (Eppendorf, 5810R) and 4 °C in a concentrator (Millipore, UFC903024) and exchanged with DPBS more than 500 times, finally concentrated to the appropriate volume. The concentration of the purified anti-CD73 antibody was determined by UV absorbance at 280 nm (NanoDrop). Antibody purity was determined by SEC-HPLC and SDS-PAGE. The recombinant antibody PR000506 was successfully expressed and purified for characterization.

[0499] Example 3 Antibody Engineering

[0500] The VH and VL sequences of the anti-CD73 antibody PR000506 were further optimized through germlineation and PTM removal.

[0501] In the germlineation process, the antibody VH or VL sequence is first aligned to the closest human germline sequence using an algorithm such as NCBI / Ig-BLAST. Then, residues in the frame region that differ from the germline sequence are inverted to their corresponding residues in the germline sequence. Finally, mature molecular biology techniques are used to recombine the germlined antibody composed of sequence variants.

[0502] Post-translational modifications (PTMs) are widely observed in proteins expressed in mammalian cells. Besides conserved PTM sites in antibodies (e.g., the conserved N-glycosylation site on the CH2 domain of the IgG1 antibody), other PTM sites occurring within the antibody-antigen binding site (i.e., the CDR region) can reduce antigen-binding activity or decrease chemical stability. For example, deamidation or isomerization can make molecules unstable and heterogeneous. To reduce sequence instability, PTM motifs can be removed through mutation. PTM motifs, such as isomerization motifs (e.g., DG), are identified in VH or VL sequences. "Hotspot" residues (e.g., the D or G in the DG motif) are then mutated to corresponding residues in germline sequences or other residues with similar biophysical properties. Antibodies composed of PTM-removed sequence variants are then recombined using sophisticated molecular biology techniques.

[0503] Table 2 shows the sequence analysis results for PR000506, PR000497, and PR001408. Table 3 shows the variants of PR000506 designed through germlineation and PTM removal. The amino acid sequences of the anti-CD73 antibodies are shown in Table 4, where PR000815-PR000820, PR000822, PR000824, PR000825, and PR003832-PR003836 are PTM-removed antibodies derived from PR000497. PR002078 is Tab 1, PR000752 is Tab 2, and PR000690 is Tab 3.

[0504] Table 2 Germline Gene Analysis of Anti-CD73 Antibody

[0505]

[0506] Table 3 PR000506 - Derivative Variant Designs

[0507]

[0508]

[0509] Table 4. SEQ ID NO of the anti-CD73 antibody in this application

[0510]

[0511]

[0512] Example 4: Binding of anti-CD73 antibody to CD73

[0513] The binding of antibodies to human CD73 and monkey CD73 expressed in stable cell lines was characterized by flow cytometry. Cells were incubated with various concentrations of antibody (maximum concentration 50 nM, 8 spots, dilution factor 3) at 4 °C for 1 h. After washing twice with PBS, secondary antibody goat anti-human IgG (H+L) (Thermo Fisher, A11013) was added, and the cells were incubated at 4 °C for 30 min. Fluorescence intensity was read using a FACS machine (BD FACS CantoII). A graph showing the relationship between the median mean fluorescence intensity of antigen-antibody binding and antibody concentration was fitted and calculated using GraphPad Prism 5, and the binding EC50 was calculated. The results are shown in Figures 1-5 and Tables 5-8.

[0514] Figure 1 and Figure 3 The binding activity of the anti-CD73 antibody with the CHO-K1-hCD73 stable cell line was shown. Tables 5 and 6 show the EC50 of the PTM-removed antibody against the CHO-K1-hCD73 stable cell line. Table 7 shows the ratio of the EC50 of the PTM-removed antibody to the EC50 of PR000506. Figure 2 The binding activity of the anti-CD73 antibody to the HEK293-cynoCD73 stable cell line was demonstrated. Table 8 shows the EC50 of the PTM-removed antibody against the HEK293-cynoCD73 stable cell line. Figure 4 Figure 5 shows the binding activity of the anti-CD73 antibody to the CHO-K1-hCD73 stable cell line. The results indicate that all antibodies have a fairly high affinity for cell-based CD73.

[0515] PR000506 PR000843 PR000844 PR000845 PR000846 PR000851 EC 50(nM) 0.7342 1.855 0.8358 0.8527 0.7499 0.7997

[0516] Tab1 PR000506 PR000847 PR000848 PR000849 PR000850 EC 50(nM) 2.172 1.047 3.026 1.528 1.079 0.8790

[0517] antibody name Antibody EC50 / PR000506 EC50 Tab1 2.074 PR000843 2.527 PR000844 1.138 PR000845 1.161 PR000846 1.021 PR000847 2.890 PR000848 1.459 PR000849 1.031 PR000850 0.840 PR000851 1.089

[0518] Tab1 PR000506 PR000844 PR000846 PR000849 PR000850 EC 50(nM) 0.6229 0.3225 0.4750 0.3071 0.5146 0.3467

[0519] Example 5: Inhibitory effect of anti-CD73 antibody on 5′ exonuclease

[0520] 5.1 Anti-CD73 antibody inhibits cell-based CD73 enzyme activity

[0521] The functional activity of anti-CD73 antibodies was determined in vitro by measuring the degree of inhibition of CD73-catalyzed AMP hydrolysis to adenosine in a stable CHO-K1-hCD73 cell line. Cells were digested with trypsin-EDTA, centrifuged at 1000 rpm for 5 minutes, and resuspended in serum-free F12K medium. Cell counting was performed using a Thermofisher, Countess... TM Cells were counted using an automated cell counter (II), and then seeded at a concentration of 5000 cells per 100 μL into 96-well plates (corning, 3599). A 4-fold diluted antibody (8 concentration points, 1:5 diluted with serum-free medium) was added to the plates, and the plates were incubated at 37°C for 30 minutes. Then, 50 μL of 4×AMP (final concentration 200 μM) was added to the plates, and the plates were incubated at 37°C for 1 hour. The plates were centrifuged at 1500 rpm for 3 minutes, and 50 μL of culture supernatant was transferred to black 96-well plates (PerkinElmer, 6005225), followed by the addition of 2×ATP (final concentration 65 μM). 100 μL / well of Cell Titer-Glo (PROMEGA, G7573) was added according to the manufacturer's instructions, and the luminescence values ​​were recorded using a PE Enspire workstation. The blocking activity is calculated using the following formula: Blocking% = [(Maximum luminescence value - Sample luminescence value) / (Maximum luminescence value - Minimum luminescence value)] × 100.

[0522] Note: AMP, ATP, and cells were added to the largest luminescent well. AMP and ATP were added to the smallest luminescent well, but no cells were present.

[0523] An EC 50 plot was created using GraphPad Prism analysis. The results are shown in Figure 6. Figures 6A-6B The inhibition of CD73 enzyme activity on the surface of CHO-K1 cells by the anti-CD73 antibody was demonstrated. Tables 9 and 10 show the EC50 of antibody binding to the CHO-K1-hCD73 stable cell line. It can be seen that all antibodies have the ability to block CD73 enzyme activity on the cell surface. Furthermore, PR000506 exhibits superior blocking activity compared to Tab1.

[0524] Tab1 PR000506 PR000844 PR000846 EC50(nM) 0.1218 0.07376 0.2073 0.1514

[0525] Tab1 PR000849 PR000850 EC50(nM) 0.07330 0.2243 0.1091

[0526] 5.2 Anti-CD73 antibody inhibits the activity of soluble CD73 enzyme.

[0527] Soluble recombinant CD73 activity was measured using the malachite green assay. 12.5 μl of 1 nM recombinant CD73 protein prepared with experimental buffer and 12.5 μl of 1 nM antibody prepared with experimental buffer were added to a 384-well plate (Corning, 3799). The assay buffer consisted of 25 mM Tris pH 7.5, 5 mM MgCl2, and 0.005% Tween-20. The plate was incubated at room temperature for 1 hour. 25 μl of AMP (maximum concentration 200 μM, diluted 1:2 with assay buffer for 8 concentration points) was added, and the samples were incubated at room temperature for 15 minutes. The concentration of inorganic phosphate was determined using the malachite green assay according to the manufacturer's instructions. The optical density at 620 nm was recorded using a PE Envision workstation. Graphs were plotted using GraphPad Prism. The results are shown in Figures 7 and 8. It can be seen that all antibodies blocked the enzymatic activity of soluble CD73 and inhibited the ability of CD73 to deadenosine from AMP.

[0528] Example 6: Internalization of CD73 by anti-CD73 antibody

[0529] 6.1 Flow cytometry detection of antibody-mediated CD73 internalization assay

[0530] NCI-H292 cells were incubated at 37°C for 4 hours or 30 minutes in the presence of 50 nM antibody or negative control isotype IgG1. Plates were washed twice with PBS, and then 100 μl of 50 nM antibody was added to the IgG1 wells for isotype control. The plates were incubated at 4°C for 1 hour, and washed twice with PBS. The wash buffer was aspirated, and 100 μl of FACS buffer containing secondary antibody goat anti-human IgG (H+L) (Thermofisher, A-21445) was added to each well. The plates were incubated at 4°C for 30 minutes. The plates were washed twice with PBS, and the cells were resuspended in 100 μl of PBS. The median fluorescence intensity was then read using a FACS Machine (BD FACS CantoII), and the surface residual % was calculated using the following formula: [Antibody 4h / (Isotype 4h + Antibody)] × 100.

[0531] The results show Figure 9A and Figure 11 The antibody in this application can mediate CD73 internalization after 4 hours.

[0532] 6.2 Antibody-mediated CD73 internalization assay using an internalization kit

[0533] CHO-K1-hCD73 cells were digested with trypsin-EDTA, then centrifuged at 1000 rpm for 5 minutes and resuspended in F12K medium containing 10% FBS. Cells were counted using a Thermofisher Countess™ II automated cell counter and seeded at 90 μl per 9000 cells per 96-well TC plate (PerkinElmer, 6005225). The plates were incubated overnight at 37°C with 5% CO2. A 10× diluted antibody (maximum concentration 10 nM, 6 concentration points, 1:5 dilution with complete F12K medium) was added to the plates to prepare a 50 μg / ml Hu-zap buffer (50×). 2 μl of this buffer was added to each well, resulting in a final concentration of 1 μg / ml. The plates were incubated at 37°C with 5% CO2 for 3 days. Add 100 μL / well of Cell Titer-Glo (PROMEGA, G7573) according to the manufacturer's instructions. Record luminescence using a PE Enspire workstation. Results are calculated as CD73 internalization % . Note: Control wells were simply seeded with cells and then topped up with culture medium to the same volume as antibody wells. Graphs were generated using GraphPad Prism.

[0534] The results show Figure 9B and Figure 10 The antibody described in this application can mediate CD73 internalization and inhibit cell growth after 3 days.

[0535] Example 7: Antibody Inhibition of AMP-Mediated CD4+ T Cell Proliferation Assay

[0536] 7.1 Isolation of CD4+ T cells from fresh human PBMCs

[0537] PBMCs were washed twice with pre-chilled PBS by centrifugation at 300×g for 10 minutes at 4°C. The supernatant was discarded, and the cells were resuspended in 20 ml of pre-chilled PBS. Cell count and viability were determined using a cell counter, followed by centrifugation at 300×g for 7 minutes. The supernatant was completely discarded. CD4+ T cells were isolated according to the manufacturer's protocol (Miltenyi, 130-045-101).

[0538] 7.2 CFSE Marking

[0539] CellTrace is ready to use and can be configured immediately. TM The stock solution (Invitrogen, C34554) was prepared by injecting it into a small vial of CellTrace. TMAdd an appropriate amount of DMSO (component B) to reagent (component A) and mix thoroughly. Dilute CellTrace in preheated (37°C) phosphate-buffered saline (PBS) or other protein-free buffer. TM Label cells with DMSO stock solution (5 mM) to 1 μM (5000 times) according to the manufacturer's instructions.

[0540] 7.3 T cell activation and expansion

[0541] CD4+ T cells from each donor were washed twice with warm, complete culture medium and diluted at 2 × 10⁻⁶ ml. 6 The amount of cells in suspension. Immunomagnetic beads containing human T cell activator CD3 / CD28 (life, 11131D) at 1×10 6 / ml suspension. Mix cells and immunomagnetic beads at a 1:1 ratio. Add 100 μl / well of the cell and bead mixture to a 96-well flat plate.

[0542] 7.4 Add 4× antibody

[0543] Add 50 μl of 4× antibody. Antibody concentration: 6 spots (maximum concentration is 20 nM, diluted 1 / 10 with RPMI 1640 complete medium for 6 spots). Incubate the plates at 37°C and 5% CO2 for 30 minutes.

[0544] 7.5 Add 4×AMP

[0545] Add 50 μL of 4×AMP (Sigma) (final concentration of 650 μM or 800 μM) and incubate the plate at 37 °C and 5% CO2 for 96 h.

[0546] 7.6 Flow Cytometry Analysis

[0547] Centrifuge at 350g for 5 minutes to precipitate the cells, wash once with 200μl PBS, and finally resuspend the cells in 200μl PBS for flow cytometry analysis on BD FACS Canto II.

[0548] The results are shown in Figures 12 and 13. The antibody can relieve AMP-mediated CD4 activation. + Inhibition of T cell proliferation. The inhibitory effect is dose-dependent.

[0549] Example 8 PK Analysis

[0550] 8.1 Flatbed Packaging

[0551] Dilute the CD73 antigen to 5 μg / mL with PBS. Add 50 μL / well to an ELISA plate. Seal the plate and incubate overnight at 4°C. Wash the plate three times with 300 μL of wash buffer. Add 200 μL / well of blocking buffer and incubate at room temperature for 1–3 hours. Wash the plate three times with 300 μL of wash buffer.

[0552] 8.2 Preparation of standards, quality control samples, test samples, and sample incubation

[0553] Standards: Standards were diluted 1:2 with 5% mixed mouse serum. The concentration range of the eight standards was 50 to 0.391 ng / mL. The sensitivity was 0.391 ng / mL. Quality Control Preparation: Two sets of quality controls were prepared in 5% mixed mouse serum: HQC (40 ng / mL), MQC (10 ng / mL), and LQC (2 ng / mL). Sample Preparation: All serum samples were first diluted 20-fold in experimental diluent. Further dilutions were performed in 5% mixed mouse serum. 50 μL of standard solution, quality controls, and samples were added to an ELISA plate. The plate was sealed and incubated at 37°C for approximately 1 hour.

[0554] 8.3 Detection reagent reaction

[0555] Wash the plate three times with 300 μl of washing buffer. Prepare the working solution of the HRP-binding detection antibody: dilute the antibody solution 1:5000 in experimental diluent. Add 100 μl of the working antibody solution to each well of the assay plate. Seal the plate and incubate at 37°C for 30 minutes.

[0556] 8.4 Substrate reaction and plate reading

[0557] Wash the plate three times with 300 μl of washing buffer. Prepare the TMB working solution: allow the substrate to stand at room temperature for 30 minutes before use. Mix equal volumes of substrate A and substrate B. Add 100 μl / well of the premixed TMB substrate and incubate at room temperature for approximately 6 minutes. Add 100 μL / well of ELISA stop solution. Mix well and read the values ​​using a SpectraMax M2 at 450 / 630 nm wavelengths. Perform data processing using SoftMax Pro GxP. Fit the standard curve using a 4-PL model with a coefficient of 1 / y.

[0558] Figure 14A Table 11 shows the antibody concentration-time curves (N = 6 / time point) of PR000506 in female C57BL / 6 mice after administration of an IV dose of 5 mg / kg. Figure 14BTable 12 shows the antibody concentration-time curves (N = 3 / time point) of PR000846 in female C57BL / 6 mice after IV administration of 10 mg / kg. The serum t-time of PR000846 is also shown. 1 / 2 The timeframe is 17.4 days for PR000846 and 18.1 days for PR000846. Figure 14C Table 13 shows the antibody concentration-time curves (N = 6 / time point) of PR000497 after IV administration of 5 mg / kg to female C57BL / 6 mice.

[0559] Table 11. 5 mg / kg dose, female C57BL / 6 mice (N=6 / time point) PR000506

[0560]

[0561]

[0562] PK parameters unit mean SD CV (%) CL ml / day / kg 3.23 0.454 14.4 Vss ml / kg 81.7 9.37 9.04 <![CDATA[Terminal t 1 / 2 ]]> day 18.1 1.54 12.9 <![CDATA[AUC last ]]> day*μg / ml 1329 137 14.3 <![CDATA[AUC INF ]]> day*μg / ml 3098 430 15.5

[0563] PK parameters unit mean SD CV (%) CL ml / day / kg 4.83 0.283 5.87 Vss ml / kg 80.7 4.59 5.70 <![CDATA[Terminal t 1 / 2 ]]> day 12.3 1.09 8.89 <![CDATA[AUC last ]]> day*μg / ml 591 21.0 3.55 <![CDATA[AUC INF ]]> day*μg / ml 1039 60.2 5.79

[0564] Example 9: In vivo tumor suppression

[0565] On day 0, tumor models were obtained by subcutaneously inoculating NCG mice with NCI-H292 cells and PBMCs. Tumor-bearing mice were divided into the following groups: isotype control group, tab1 group, PR000506 group, tab3 group, PR000846 group, and PR000497 group. In each group, mice were treated with a 10 mpk dose of antibody twice a week for three consecutive weeks. Tumor volume and mouse body weight were measured at selected time points.

[0566] The results are shown in Figures 15 and 16. It can be seen that the anti-CD73 antibody has an inhibitory effect on tumor growth.

[0567] Example 10: Determination of the epitope of CD73 bound by PR000846

[0568] In this embodiment, the epitope of the anti-CD73 antibody PR000846 binding to CD73 was determined by hydrogen-deuterium exchange mass spectrometry (HDX-MS), and the key amino acid of the epitope was confirmed by scanning the alanine mutation of the CD73 recombinant protein.

[0569] Hydrogen-deuterium exchange mass spectrometry (HDX-MS)

[0570] HDX-MS experiments were performed using Genechem, as described in the following literature (Park IH et al., J. Chem. Inf. Model.; 55(9):1914-1925 (2015); Chalmers MJ et al., Anal. Chem.; 78(4):1005-14 (2006)). Antibody-antigen complexes were prepared by overnight incubation at 4°C using a 1:1 molar ratio.

[0571] Take 2 μl of 0.5 mg / ml recombinant human CD73 protein (Novoprotein, C446), add 2 μl of PBS or 2 μl of an equimolar amount of the test antibody, and then add to 36 μl of D2O buffer. Perform exchanges at 30 s, 120 s, 600 s, 1800 s, and 7200 s at room temperature, then add 40 μl of pre-chilled Quench Buffer (0.5 M TCEP, 4 M guanidine hydrochloride, pH = 2.5) at 4 °C and mix well to stop the reaction. After digestion with Pepsin online enzyme column, hydrogen-deuterium exchange data were acquired using LC-MS. Chromatographic gradient and coverage tests were performed. Hydrogen-deuterium exchange data analysis: Peptide libraries were constructed using ProteomeDiscoverer 2.1, and hydrogen-deuterium exchange data were analyzed using HDExaminer.

[0572] Figure 17-20 The results of hydrogen-deuterium exchange MS (HDX-MS) analysis are described, showing the differences in deuterium uptake values ​​of various peptides in the hydrogen-deuterium exchange experiments of CD73 and CD73+PR000846.

[0573] Figure 17 Each data point indicates the difference in deuterium uptake between the CD73+PR000846 complex (positive values ​​on the y-axis) and CD73 alone (negative values ​​on the y-axis). The vertical bars represent the sum of uptake differences over the entire exposure time-point. The horizontal axis corresponds to the peptides analyzed from the N- to C-terminus (from left to right). Comparing the exchange kinetics between free and complexed CD73 revealed that when PR000846 is bound, three regions within the N-terminal domain of sCD73 (region 1: aa115–164; region 2: aa168–191; region 3: aa222–254) reduced deuterium uptake.

[0574] Figure 18-20As shown, the relative deuterium uptake (Dalton mass change) in peptides in regions 1, 2, and 3 changes at different time points. To narrow the sequence regions containing hydrogen exchange changes so as to predict epitopes, the relative mass changes of overlapping peptides were compared. For example, in region 2, the peptide spanning positions 168-186 shows differential deuterium exchange, while the peptide spanning positions 168-181 shows no difference. Therefore, it can be inferred that the residues upstream of position 182 are not differentially labeled with deuterium. Thus, it can be inferred that the three peptide regions of PR000846 binding to CD73 can be narrowed down to (amino acids (aa) 132-146 (YLPYKVGDEVVG), 182-189 (KTLNVNKI), and 240-254 (IVTSDDGRKVPVVQA)), showing reduced deuterium uptake.

[0575] Example 11 Alanine point mutation in CD73

[0576] To obtain a more accurate epitope map, alanine point mutations were performed on these three regions of CD73. Plasmids were constructed by performing alanine point mutations on these three regions of CD73, transfected into HEK293 cells, and expressed. Then, ELISA was used to detect the binding of the CD73 alanine point mutant to the antibody, confirming the key amino acids of the antigen-binding epitope, as follows: The antibody was coated with 0.5 μg / ml anti-His antibody (Vendor TransGen, HT501-02), blocked with 3% milk PBS at 37°C for 2 hours, 100 μL of CD73 mutant supernatant was added, and incubated at 37°C for 30 minutes. This process was repeated three times to saturate the CD73 mutant supernatant. Then, serially diluted PR000846 (initial concentration 100 nM) was added at 37°C for 1 hour. Anti-CD73 antibody and 0.5% blocking buffer were added at a ratio of 1:5000, and incubated at 37°C for 1 hour. The sample was washed three times with PBST (0.05% Tween 20), 100 μL of TMB (KPL) was added, and incubated at room temperature for approximately 5 minutes. Finally, 50 μL of stop solution was added. The absorbance (density at 450 nm) of each well was read using a plate reader.

[0577] Figure 21 and 22ELISA combined data show that K136, V137, L138, D142, E143, V144 in region 1, K180, V186, N187, K188 in region 2, and Y223, P238, I240 in or near region 3 are the major epitope residues for CD73 binding to PR000846. Furthermore, V137 and D142 in region 1 are key epitope residues. N185 is a key epitope residue for CD73 binding to MEDI9447(TAB1), but not for CD73 binding to PR000846.

[0578] Although preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. The invention should not be limited to the specific instances provided in the specification. Although the invention has been described with reference to the foregoing description, the description and illustrations of the embodiments herein are not intended to be construed as limiting. Many variations, alterations, and substitutions will now occur to those skilled in the art without departing from the invention. Furthermore, it should be understood that all aspects of the invention are not limited to the specific descriptions, constructions, or relative proportions set forth herein, which depend on various conditions and variables. It should be understood that various alternatives to the embodiments of the invention described herein can be used to implement the invention. Therefore, it is contemplated that the invention will also cover any such alternatives, modifications, variations, or equivalents. The following claims define the scope of the invention and thereby cover the methods and structures within the scope of these claims and their equivalents. sequence list <110> Harbour BioMed (Shanghai) Co., Ltd. <120> Anti-CD73 antibodies and their applications <130> 0113-PA-013CN.DIV1 <160> 169 <170> PatentIn version 3.5 <210> 1 <211> 25 <212> PRT <213> Artificial sequence <220> <223> PR000497 HFWR1 Chothia, PR000815 HFWR1 Chothia, PR000816 HFWR1 Chothia <400> 1 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser 20 25 <210> 2 <211> 25 <212> PRT <213> artificial sequence <220> <223> PR000506 HFWR1 Chothia, PR000843 HFWR1 Chothia, PR000844 HFWR1Chothia <400> 2 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser 20 25 <210> 3 <211> 25 <212> PRT <213> artificial sequence <220> <223> PR000690 HFWR1 Chothia, PR000817 HFWR1 Chothia, PR000818 HFWR1Chothia <400> 3 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser 20 25 <210> 4 <211> 25 <212> PRT <213> artificial sequence <220> <223> PR000752 HFWR1 Chothia <400> 4 Glu Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser 20 25 <210> 5 <211> 25 <212> PRT <213> Artificial sequence <220> <223> PR002078 HFWR1 Chothia <400> 5 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser 20 25 <210> 6 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000497 HCDR1 Chothia, PR000815 HCDR1 Chothia, PR000816 HCDR1 Chothia <400> 6 Gly Phe Thr Phe Ser Ser Tyr 1 5 <210> 7 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000506 HCDR1 Chothia, PR000843 HCDR1 Chothia, PR000844 HCDR1 Chothia <400> 7 Gly Phe Thr Phe Ser Asp Tyr 1 5 <210> 8 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000690 HCDR1 Chothia <400> 8 Gly Phe Thr Phe Ser Asn Tyr 1 5 <210> 9 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000752 HCDR1 Chothia <400> 9 Gly Tyr Ala Phe Thr Ser Tyr 1 5 <210> 10 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR001408 HCDR1 Chothia <400> 10 Gly Phe Thr Phe Tyr Ser Tyr 1 5 <210> 11 <211> 19 <212> PRT <213> Artificial sequence <220> <223> PR000497 HFWR2 Chothia, PR000815 HFWR2 Chothia, PR000816 HFWR2Chothia <400> 11 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 1 5 10 15 Ala Ser Thr <210> 12 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> PR000506 HFWR2 Chothia, PR000843 HFWR2 Chothia, PR000844 HFWR2Chothia <400> 12 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 1 5 10 15 Ala Leu Ile <210> 13 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> PR000690 HFWR2 Chothia <400> 13 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 1 5 10 15 Ala Val Ile <210> 14 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> PR000752 HFWR2 Chothia <400> 14 Asn Met Tyr Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 1 5 10 15 Gly Tyr Ile <210> 15 <211> 19 <212> PRT <213> Artificial sequence <220> <223> PR001408 HFWR2 Chothia <400> 15 Gly Met His Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu Trp Val 1 5 10 15 Ala Leu Ile <210> 16 <211> 19 <212> PRT <213> Artificial sequence <220> <223> PR002078 HFWR2 Chothia <400> 16 Ala Tyr Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 1 5 10 15 Ser Ala Ile <210> 17 <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000497 HCDR2 Chothia, PR000819 HCDR2 Chothia, PR000820 HCDR2 Chothia <400> 17 Trp Tyr Asp Gly Ser Lys 1 5 <210> 18 <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000506 HCDR2 Chothia, PR000851 HCDR2 Chothia <400> 18 Trp Tyr Asp Gly Ser Phe 1 5 <210> 19 <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000690 HCDR2 Chothia <400> 19 Leu Tyr Asp Gly Ser Asn 1 5 <210> 20 <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000752 HCDR2 Chothia <400> 20 Asp Pro Tyr Asn Gly Gly 1 5 <210> twenty one <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000815 HCDR2 Chothia, PR000817 HCDR2 Chothia <400> twenty one Trp Tyr Glu Gly Ser Lys 1 5 <210> twenty two <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000816 HCDR2 Chothia, PR000818 HCDR2 Chothia, PR000822 HCDR2 Chothia <400> twenty two Trp Tyr Asp Ala Ser Lys 1 5 <210> twenty three <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000843 HCDR2 Chothia, PR000844 HCDR2 Chothia, PR000847 HCDR2 Chothia <400> twenty three Trp Tyr Glu Gly Ser Phe 1 5 <210> twenty four <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR000845 HCDR2 Chothia, PR000846 HCDR2 Chothia, PR000849 HCDR2 Chothia <400> twenty four Trp Tyr Asp Ala Ser Phe 1 5 <210> 25 <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR001408 HCDR2 Chothia <400> 25 Trp Tyr Asp Gly Ser Asn 1 5 <210> 26 <211> 6 <212> PRT <213> Artificial sequence <220> <223> PR002078 HCDR2 Chothia <400> 26 Ser Gly Ser Gly Gly Arg 1 5 <210> 27 <211> 41 <212> PRT <213> artificial sequence <220> <223> PR000497 HFWR3 Chothia, PR000815 HFWR3 Chothia, PR000816 HFWR3Chothia <400> 27 Lys Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 1 5 10 15 Asn Ser Lys Asn Thr Leu Tyr Leu Lys Met Asn Ser Leu Arg Gly Asp 20 25 30 Asp Thr Ala Val Tyr Tyr Cys Ala Lys 35 40 <210> 28 <211> 41 <212> PRT <213> artificial sequence <220> <223> PR000506 HFWR3 Chothia, PR000843 HFWR3 Chothia, PR000844 HFWR3Chothia <400> 28 Glu Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Ser Ile Ser Arg Asp 1 5 10 15 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 20 25 30 Asp Thr Ala Val Tyr Tyr Cys Val Arg 35 40 <210> 29 <211> 41 <212> PRT <213> artificial sequence <220> <223> PR000690 HFWR3 Chothia <400> 29 Lys Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 1 5 10 15 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 20 25 30 Asp Thr Ala Val Tyr Tyr Cys Ala Arg 35 40 <210> 30 <211> 41 <212> PRT <213> artificial sequence <220> <223> PR000752 HFWR3 Chothia <400> 30 Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr Val Asp 1 5 10 15 Lys Ser Ser Ser Thr Ala Tyr Met His Leu Asn Ser Leu Thr Ser Glu 20 25 30 Asp Ser Ala Val Tyr Tyr Cys Ala Arg 35 40 <210> 31 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> PR000817 HFWR3 Chothia, PR000818 HFWR3 Chothia, PR000819 HFWR3Chothia <400> 31 Lys Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 1 5 10 15 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 20 25 30 Asp Thr Ala Val Tyr Tyr Cys Ala Lys 35 40 <210> 32 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> PR001408 HFWR3 Chothia <400> 32 Asn Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 1 5 10 15 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 20 25 30 Asp Thr Ala Val Tyr Tyr Cys Ala Thr 35 40 <210> 33 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> PR002078 HFWR3 Chothia <400> 33 Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 1 5 10 15 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 20 25 30 Asp Thr Ala Val Tyr Tyr Cys Ala Arg 35 40 <210> 34 <211> 13 <212> PRT <213> Artificial sequence <220> <223> PR000497 HCDR3 Chothia, PR000815 HCDR3 Chothia, PR000816 HCDR3 Chothia <400> 34 Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile 1 5 10 <210> 35 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000506 HCDR3 Chothia, PR000851 HCDR3 Chothia <400> 35 Asp Gly Gln Trp Gly Ser Arg Leu Asp Tyr 1 5 10 <210> 36 <211> 12 <212> PRT <213> Artificial sequence <220> <223> PR000690 HCDR3 Chothia <400> 36 Gly Gly Ser Ser Trp Tyr Pro Asp Ser Phe Asp Ile 1 5 10 <210> 37 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000752 HCDR3 Chothia <400> 37 Gly Tyr Gly Asn Tyr Lys Ala Trp Phe Ala Tyr 1 5 10 <210> 38 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000843 HCDR3 Chothia, PR000845 HCDR3 Chothia, PR000847 HCDR3 Chothia <400> 38 Glu Gly Gln Trp Gly Ser Arg Leu Asp Tyr 1 5 10 <210> 39 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000844 HCDR3 Chothia, PR000846 HCDR3 Chothia, PR000848 HCDR3 Chothia <400> 39 Asp Ala Gln Trp Gly Ser Arg Leu Asp Tyr 1 5 10 <210> 40 <211> 14 <212> PRT <213> Artificial sequence <220> <223> PR001408 HCDR3 Chothia <400> 40 Glu Arg Ser Ser Ser Phe Tyr Tyr Tyr Tyr Gly Met Asp Val 1 5 10 <210> 41 <211> 8 <212> PRT <213> Artificial sequence <220> <223> PR002078 HCDR3 Chothia <400> 41 Leu Gly Tyr Gly Arg Val Asp Glu 1 5 <210> 42 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000497 HFWR4 Chothia, PR000690 HFWR4 Chothia, PR000815 HFWR4Chothia <400> 42 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 1 5 10 <210> 43 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000506 HFWR4 Chothia, PR000843 HFWR4 Chothia, PR000844 HFWR4 Chothia <400> 43 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 1 5 10 <210> 44 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000752 HFWR4 Chothia <400> 44 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 1 5 10 <210> 45 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR001408 HFWR4 Chothia <400> 45 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 1 5 10 <210> 46 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR002078 HFWR4 Chothia <400> 46 Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 1 5 10 <210> 47 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> PR000497 LFWR1 Chothia, PR000815 LFWR1 Chothia, PR000816 LFWR1 Chothia <400> 47 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys 20 <210> 48 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> PR000506 LFWR1 Chothia, PR000843 LFWR1 Chothia, PR000844 LFWR1 Chothia <400> 48 Lys Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys 20 <210> 49 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> PR000690 LFWR1 Chothia <400> 49 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys 20 <210> 50 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> PR000752 LFWR1 Chothia <400> 50 Asp Ala Val Met Thr Gln Thr Pro Lys Phe Leu Leu Val Ser Ala Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys 20 <210> 51 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> PR000847 LFWR1 Chothia, PR000848 LFWR1 Chothia, PR000849 LFWR1 Chothia <400> 51 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys 20 <210> 52 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> PR001408 LFWR1 Chothia <400> 52 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys 20 <210> 53 <211> twenty two <212> PRT <213> Artificial sequence <220> <223> PR002078 LFWR1 Chothia <400> 53 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys 20 <210> 54 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000497 LCDR1 Chothia, PR000815 LCDR1 Chothia, PR000816 LCDR1Chothia <400> 54 Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala 1 5 10 <210> 55 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000506 LCDR1 Chothia, PR000843 LCDR1 Chothia, PR000844 LCDR1Chothia <400> 55 Arg Ala Ser Gln Ser Val Ser Arg Tyr Leu Ala 1 5 10 <210> 56 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000690 LCDR1 Chothia <400> 56 Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala 1 5 10 <210> 57 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR000752 LCDR1 Chothia <400> 57 Lys Ala Ser Gln Ser Val Thr Asn Asp Val Ala 1 5 10 <210> 58 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR001408 LCDR1 Chothia <400> 58 Arg Ala Ser Gln Gly Ile Ser Ser Tyr Leu Ala 1 5 10 <210> 59 <211> 13 <212> PRT <213> Artificial sequence <220> <223> PR002078 LCDR1 Chothia <400> 59 Ser Gly Ser Leu Ser Asn Ile Gly Arg Asn Pro Val Asn 1 5 10 <210> 60 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PR000497 LFWR2 Chothia, PR000815 LFWR2 Chothia, PR000816 LFWR2 Chothia <400> 60 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 1 5 10 15 <210> 61 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PR000506 LFWR2 Chothia, PR000843 LFWR2 Chothia, PR000844 LFWR2 Chothia <400> 61 Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 1 5 10 15 <210> 62 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PR000690 LFWR2 Chothia <400> 62 Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile Tyr 1 5 10 15 <210> 63 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PR000752 LFWR2 Chothia <400> 63 Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr 1 5 10 15 <210> 64 <211> 15 <212> PRT <213> Artificial sequence <220> <223> PR002078 LFWR2 Chothia <400> 64 Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr 1 5 10 15 <210> 65 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000497 LCDR2 Chothia, PR000815 LCDR2 Chothia, PR000816 LCDR2Chothia <400> 65 Lys Ala Ser Ser Leu Glu Ser 1 5 <210> 66 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000506 LCDR2 Chothia, PR000843 LCDR2 Chothia, PR000844 LCDR2Chothia <400> 66 Asp Ala Ser Asn Arg Ala Thr 1 5 <210> 67 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000690 LCDR2 Chothia <400> 67 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 68 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR000752 LCDR2 Chothia <400> 68 Tyr Ala Ser Asn Arg Tyr Thr 1 5 <210> 69 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR001408 LCDR2 Chothia <400> 69 Ala Ala Ser Thr Leu Gln Ser 1 5 <210> 70 <211> 7 <212> PRT <213> Artificial sequence <220> <223> PR002078 LCDR2 Chothia <400> 70 Leu Asp Asn Leu Arg Leu Ser 1 5 <210> 71 <211> 32 <212> PRT <213> Artificial sequence <220> <223> PR000497 LFWR3 Chothia, PR000815 LFWR3 Chothia, PR000816 LFWR3 Chothia <400> 71 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr 1 5 10 15 Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys 20 25 30 <210> 72 <211> 32 <212> PRT <213> Artificial sequence <220> <223> PR000506 LFWR3 Chothia, PR000843 LFWR3 Chothia, PR000844 LFWR3 Chothia <400> 72 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 20 25 30 <210> 73 <211> 32 <212> PRT <213> Artificial sequence <220> <223> PR000690 LFWR3 Chothia <400> 73 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 20 25 30 <210> 74 <211> 32 <212> PRT <213> Artificial sequence <220> <223> PR000752 LFWR3 Chothia <400> 74 Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Tyr Gly Thr Asp Phe Thr 1 5 10 15 Phe Thr Ile Ser Thr Val Gln Ala Glu Asp Leu Ala Val Tyr Phe Cys 20 25 30 <210> 75 <211> 32 <212> PRT <213> Artificial sequence <220> <223> PR001408 LFWR3 Chothia <400> 75 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr 1 5 10 15 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 20 25 30 <210> 76 <211> 32 <212> PRT <213> Artificial sequence <220> <223> PR002078 LFWR3 Chothia <400> 76 Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser 1 5 10 15 Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys 20 25 30 <210> 77 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR000497 LCDR3 Chothia, PR000815 LCDR3 Chothia, PR000816 LCDR3Chothia <400> 77 Gln Gln Tyr Asp Asn Tyr Ser Asn Thr 1 5 <210> 78 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR000506 LCDR3 Chothia, PR000843 LCDR3 Chothia, PR000844 LCDR3Chothia <400> 78 Gln Gln Arg Ser Asn Trp Ile Phe Thr 1 5 <210> 79 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR000690 LCDR3 Chothia <400> 79 Gln Gln Tyr Asn Ser Tyr Pro Leu Thr 1 5 <210> 80 <211> 8 <212> PRT <213> Artificial sequence <220> <223> PR000752 LCDR3 Chothia <400> 80 Gln Gln Asp Tyr Ser Ser Leu Thr 1 5 <210> 81 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR000820 LCDR3 Chothia, PR000822 LCDR3 Chothia, PR000824 LCDR3Chothia <400> 81 Gln Gln Tyr Asp Gln Tyr Ser Asn Thr 1 5 <210> 82 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR001408 LCDR3 Chothia <400> 82 Gln Gln Leu Asn Ser Tyr Pro Tyr Thr 1 5 <210> 83 <211> 11 <212> PRT <213> Artificial sequence <220> <223> PR002078 LCDR3 Chothia <400> 83 Ala Thr Trp Asp Asp Ser His Pro Gly Trp Thr 1 5 10 <210> 84 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR003832 LCDR3 Chothia <400> 84 Gln Gln Tyr Asp Met Tyr Ser Asn Thr 1 5 <210> 85 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR003833 LCDR3 Chothia <400> 85 Gln Gln Tyr Asp Thr Tyr Ser Asn Thr 1 5 <210> 86 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR003834 LCDR3 Chothia <400> 86 Gln Gln Tyr Asp Ser Tyr Ser Asn Thr 1 5 <210> 87 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR003835 LCDR3 Chothia <400> 87 Gln Gln Tyr Asp Asn Tyr Lys Asn Thr 1 5 <210> 88 <211> 9 <212> PRT <213> Artificial sequence <220> <223> PR003836 LCDR3 Chothia <400> 88 Gln Gln Tyr Asp Asn Tyr Glu Asn Thr 1 5 <210> 89 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000497 LFWR4 Chothia, PR000815 LFWR4 Chothia, PR000816 LFWR4 Chothia <400> 89 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 1 5 10 <210> 90 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000506 LFWR4 Chothia, PR000843 LFWR4 Chothia, PR000844 LFWR4 Chothia <400> 90 Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 1 5 10 <210> 91 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000690 LFWR4 Chothia <400> 91 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 1 5 10 <210> 92 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR000752 LFWR4 Chothia <400> 92 Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 1 5 10 <210> 93 <211> 10 <212> PRT <213> Artificial sequence <220> <223> PR001408 LFWR4 Chothia <400> 93 Phe Gly Gln Gly Thr Glu Leu Glu Ile Lys 1 5 10 <210> 94 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> PR002078 LFWR4 Chothia <400> 94 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 1 5 10 <210> 95 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> PR000497 VH, PR000820 VH <400> 95 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Lys Met Asn Ser Leu Arg Gly Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 96 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> PR000506 VH, PR000851 VH <400> 96 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Gly Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Asp Gly Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 97 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> PR000690 VH <400> 97 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Ala Val Ile Leu Tyr Asp Gly Ser Asn Lys Tyr Tyr Pro Asp Ser Val 35 40 45 Ala Val Ile Leu Tyr Asp Gly Ser Asn Lys Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Ser Ser Trp Tyr Pro Asp Ser Phe Asp Ile Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 98 <211> 120 <212> PRT <213> artificial sequence <220> <223> PR000752 VH <400> 98 Glu Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Ser Tyr 20 25 30 Asn Met Tyr Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asp Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met His Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Tyr Gly Asn Tyr Lys Ala Trp Phe Ala Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 99 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> PR000815 VH <400> 99 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Glu Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Lys Met Asn Ser Leu Arg Gly Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 100 <211> 122 <212> PRT <213> artificial sequence <220> <223> PR000816 VH, PR000822 VH <400> 100 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Ala Ser Light Light Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Lys Met Asn Ser Leu Arg Gly Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 101 <211> 122 <212> PRT <213> artificial sequence <220> <223> PR000817 VH <400> 101 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Glu Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 102 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> PR000818 VH, PR000824 VH, PR003832 VH, PR003833 VH, PR003834 VH, PR003 <400> 102 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Ala Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 103 <211> 122 <212> PRT <213> Artificial sequence <220> <223> PR000819 VH, PR000825 VH <400> 103 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 104 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> PR000843 VH, PR000847 VH <400> 104 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Glu Gly Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 105 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> PR000844 VH, PR000848 VH <400> 105 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Glu Gly Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Asp Ala Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 106 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> PR000845 VH, PR000849 VH <400> 106 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Ala Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 107 <211> 119 <212> PRT <213> artificial sequence <220> <223> PR000846 VH, PR000850 VH <400> 107 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Ala Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Asp Ala Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 108 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> PR001408 VH <400> 108 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Gly Ser Asn Asn Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Glu Arg Ser Ser Ser Phe Tyr Tyr Tyr Tyr Gly Met Asp Val 100 105 110 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 109 <211> 117 <212> PRT <213> artificial sequence <220> <223> PR002078 VH <400> 109 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Tyr Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Gly Tyr Gly Arg Val Asp Glu Trp Gly Arg Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 110 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR000497 VL, PR000815 VL, PR000816 VL, PR000817 VL, PR000818 VL,PR000 <400> 110 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Asn Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 111 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR000506 VL, PR000843 VL, PR000844 VL, PR000845 VL, PR000846 VL <400> 111 Lys Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Ile Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 112 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR000690 VL <400> 112 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 113 <211> 106 <212> PRT <213> artificial sequence <220> <223> PR000752 VL <400> 113 Asp Ala Val Met Thr Gln Thr Pro Lys Phe Leu Leu Val Ser Ala Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Thr Asn Asp 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Thr Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln Asp Tyr Ser Ser Leu Thr 85 90 95 Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 114 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR000820 VL, PR000822 VL, PR000824 VL, PR000825 VL <400> 114 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Gln Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 115 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR000847 VL, PR000848 VL, PR000849 VL, PR000850 VL, PR000851 VL <400> 115 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Ile Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 116 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR001408 VL <400> 116 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Glu Leu Glu Ile Lys 100 105 <210> 117 <211> 110 <212> PRT <213> Artificial Sequence <220> <223> PR002078 VL <400> 117 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Leu Ser Asn Ile Gly Arg Asn 20 25 30 Pro Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Leu Asp Asn Leu Arg Leu Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Trp Asp Asp Ser His 85 90 95 Pro Gly Trp Thr Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 <210> 118 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> PR003832 VL <400> 118 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Met Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 119 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR003833 VL <400> 119 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 120 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR003834 VL <400> 120 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Ser Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 121 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR003835 VL <400> 121 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Asn Tyr Lys Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 122 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR003836 VL <400> 122 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Asn Tyr Glu Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 123 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> PR000497 HC, PR000820 HC <400> 123 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Lys Met Asn Ser Leu Arg Gly Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 124 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> PR000506 HC, PR000851 HC <400> 124 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Gly Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Asp Gly Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 125 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> PR000690 HC <400> 125 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Leu Tyr Asp Gly Ser Asn Lys Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Ser Ser Trp Tyr Pro Asp Ser Phe Asp Ile Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly 225 230 235 240 Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 126 <211> 450 <212> PRT <213> Artificial Sequence <220> <223> PR000752 HC <400> 126 Glu Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Ser Tyr 20 25 30 Asn Met Tyr Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asp Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met His Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Tyr Gly Asn Tyr Lys Ala Trp Phe Ala Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Gln Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> 127 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> PR000815 HC <400> 127 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Glu Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Lys Met Asn Ser Leu Arg Gly Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 128 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> PR000816 HC, PR000822 HC <400> 128 Gln Val Gln Val Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Ala Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Lys Met Asn Ser Leu Arg Gly Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 129 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> PR000817 HC <400> 129 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Glu Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 130 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> PR000818 HC, PR000824 HC, PR003832 HC, PR003833 HC, PR003834 HC, PR003 <400> 130 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Ala Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 131 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> PR000819 HC, PR000825 HC <400> 131 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Thr Trp Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Asp Leu Leu Thr Gly Pro Asn Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 132 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> PR000843 HC, PR000847 HC <400> 132 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Glu Gly Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 133 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> PR000844 HC, PR000848 HC <400> 133 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Glu Gly Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Asp Ala Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 134 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> PR000845 HC, PR000849 HC <400> 134 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Ala Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 135 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> PR000846 HC, PR000850 HC <400> 135 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Ala Ser Phe Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Asp Ala Gln Trp Gly Ser Arg Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 136 <211> 453 <212> PRT <213> Artificial Sequence <220> <223> PR001408 HC <400> 136 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Leu Ile Trp Tyr Asp Gly Ser Asn Asn Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Glu Arg Ser Ser Ser Phe Tyr Tyr Tyr Tyr Gly Met Asp Val 100 105 110 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 225 230 235 240 Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln 355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450 <210> 137 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> PR002078 HC <400> 137 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Tyr Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Gly Tyr Gly Arg Val Asp Glu Trp Gly Arg Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Ser Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 138 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR000497 LC, PR000815 LC, PR000816 LC, PR000817 LC, PR000818 LC, PR000 <400> 138 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Asn Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 139 <211> 214 <212> PRT <213> artificial sequence <220> <223> PR000506 LC, PR000843 LC, PR000844 LC, PR000845 LC, PR000846 LC <400> 139 Lys Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Ile Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 140 <211> 214 <212> PRT <213> artificial sequence <220> <223> PR000690 LC <400> 140 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 141 <211> 213 <212> PRT <213> Artificial sequence <220> <223> PR000752 LC <400> 141 Asp Ala Val Met Thr Gln Thr Pro Lys Phe Leu Leu Val Ser Ala Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Thr Asn Asp 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Thr Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln Asp Tyr Ser Ser Leu Thr 85 90 95 Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 <210> 142 <211> 214 <212> PRT <213> artificial sequence <220> <223> PR000820 LC, PR000822 LC, PR000824 LC, PR000825 LC <400> 142 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Gln Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 143 <211> 214 <212> PRT <213> artificial sequence <220> <223> PR000847 LC, PR000848 LC, PR000849 LC, PR000850 LC, PR000851 LC <400> 143 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Ile Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 144 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR001408 LC <400> 144 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Glu Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 145 <211> 216 <212> PRT <213> Artificial Sequence <220> <223> PR002078 LC <400> 145 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Leu Ser Asn Ile Gly Arg Asn 20 25 30 Pro Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Leu Asp Asn Leu Arg Leu Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Trp Asp Asp Ser His 85 90 95 Pro Gly Trp Thr Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 146 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR003832 LC <400> 146 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Met Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 147 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR003833 LC <400> 147 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 148 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR003834 LC <400> 148 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Ser Tyr Ser Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 149 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR003835 LC <400> 149 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Asn Tyr Lys Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 150 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> PR003836 LC <400> 150 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Asn Tyr Glu Asn 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 151 <211> 107 <212> PRT <213> artificial sequence <220> <223> PR000506; PR000843; PR000844; PR000845; PR000846; PR000847; PR000848; PR000849; PR000850; PR000851 <220> <221> misc_feature <222> (1)..(1) <223> Xaa = Glu or Lys <220> <221> misc_feature <222> (4)..(4) <223> Xaa = Leu or Met <400> 151 Xaa Ile Val Xaa Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Ar...

Claims

1. An isolated antigen-binding protein that binds to CD73, comprising HCDR1, HCDR2, and HCDR3 of the heavy chain variable region VH; and LCDR1, LCDR2, and LCDR3 of the light chain variable region VL, wherein the antigen-binding protein is an antibody or an antigen-binding fragment thereof, wherein, (1) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:77, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:17, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (2) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:77, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:21, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (3) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:77, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (4) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:81, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:17, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (5) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:81, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (6) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:84, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (7) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:85, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (8) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:86, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (9) The amino acid sequence of LCDR1 is shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is shown in SEQ ID NO:87, the amino acid sequence of HCDR1 is shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:34; or, (10) The amino acid sequence of LCDR1 is as shown in SEQ ID NO:54, the amino acid sequence of LCDR2 is as shown in SEQ ID NO:65, the amino acid sequence of LCDR3 is as shown in SEQ ID NO:88, the amino acid sequence of HCDR1 is as shown in SEQ ID NO:6, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:22, and the amino acid sequence of HCDR3 is as shown in SEQ ID NO:34; or, (11) The amino acid sequence of LCDR1 is shown in SEQ ID NO:58, the amino acid sequence of LCDR2 is shown in SEQ ID NO:69, the amino acid sequence of LCDR3 is shown in SEQ ID NO:82, the amino acid sequence of HCDR1 is shown in SEQ ID NO:10, the amino acid sequence of HCDR2 is shown in SEQ ID NO:25, and the amino acid sequence of HCDR3 is shown in SEQ ID NO:

40.

2. The isolated antigen-binding protein that binds to CD73 according to claim 1, wherein the antigen-binding fragment comprises Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, and / or di-scFv.

3. The isolated antigen-binding protein that binds to CD73 according to claim 1 or 2, wherein the antibody is derived from a chimeric antibody, a humanized antibody, or a fully human antibody.

4. The isolated antigen-binding protein that binds to CD73 according to claim 1 or 2, comprising a heavy chain variable region VH, wherein the VH comprises an amino acid sequence shown in any one of SEQ ID NO: 95, 99, 100, 101, 102, 103 and 108.

5. The isolated antigen-binding protein binding to CD73 according to claim 1 or 2, comprising an antibody heavy chain HC, said HC comprising the amino acid sequence shown in any one of SEQ ID NO: 123, 127, 128, 129, 130, 131 and 136.

6. The isolated antigen-binding protein that binds to CD73 according to claim 1 or 2, comprising a light chain variable region VL, wherein the VL comprises the amino acid sequence shown in any one of SEQ ID NO: 110, 114, 116, 118, 119, 120, 121 and 122.

7. The isolated antigen-binding protein binding to CD73 according to claim 1 or 2, comprising an antibody light chain LC, said LC comprising the amino acid sequence shown in any one of SEQ ID NO: 138, 142, 144, 146, 147, 148, 149 and 150.

8. One or more isolated nucleic acid molecules encoding the isolated antigen-binding protein that binds to CD73 as described in any one of claims 1-7.

9. One or more carriers comprising one or more isolated nucleic acid molecules as described in claim 8.

10. A cell comprising one or more isolated nucleic acid molecules as claimed in claim 8 or one or more vectors as claimed in claim 9.

11. A method for preparing the isolated antigen-binding protein of any one of claims 1-7, the method comprising culturing the cells of claim 10 under conditions that cause the isolated antigen-binding protein of any one of claims 1-7 to be expressed.

12. A pharmaceutical composition comprising, as described in any one of claims 1-7, an isolated antigen-binding protein binding to CD73, one or more isolated nucleic acid molecules as described in claim 8, one or more carriers as described in claim 9, and / or the cell as described in claim 10, and optionally a pharmaceutically acceptable carrier.

13. Use in the preparation of a medicament of any one of claims 1-7, of any one or more isolated nucleic acid molecules of claim 8, of any one or more carriers of claim 9, of any one cell of claim 10, and / or of any one pharmaceutical composition of claim 12, wherein the medicament is used to alleviate and / or treat solid tumors expressing CD73.

14. The use according to claim 13, wherein the solid tumor is lung cancer.

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