An antibody and uses thereof

Abstract

The present invention relates to antibodies that bind primarily to plasma prekallikrein, and the use of these antibodies in the manufacture of a medicament for preventing or treating a plasma kallikrein or plasma prekallikrein related disease in a subject.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to an antibody that primarily binds to plasma kallikrein precursors and its uses. Background Technology

[0002] Plasma kallikrein (PK) belongs to the serine protease family and was first discovered in mammalian plasma. It is encoded by a single gene (KLKB1) located on chromosome 4q35 and is primarily synthesized in the liver. PK is a key enzyme in the kallikrein-kinin system (KKS), acting on high-molecular-weight kininogen (KH) to activate and release small-molecule bradykinin (BK). BK then participates in biological processes such as coagulation, fibrinolysis, complement activation, and inflammation by acting on bradykinin receptors. Plasma prekallikrein (PPK) is the precursor of plasma kallikrein. Plasma kallikrein exists in large quantities in the bloodstream as prekallikrein (PPK) and is further activated by coagulation factor XIIa through the cleavage of its intrinsic Arg-IIe bond, converting it into PK.

[0003] In recent years, with more in-depth research into the genetics, molecular biology, and pharmacology of plasma kallikrein, our understanding of its physiological and pathological roles has deepened. Studies have shown that plasma kallikrein is closely related to a variety of diseases, including inflammatory diseases, tumors, cardiovascular diseases, kidney diseases, central nervous system diseases, retinopathy, and diabetic retinopathy (Costa-Neto, CME et al. Participation of kallikrein-kinin system in different pathologies. Int. Immunopharmacol. 2008, 8, 135-142). For example, hereditary angioedema (HAE) is an autosomal dominant inherited condition mainly caused by a deficiency of C1-INH in the body, which weakens its inhibitory effect on plasma kallikrein, uncontrolledly activating the KKS system, releasing vasoactive substances, increasing vascular permeability, and causing typical swelling (Farkas, H. Orphan drugs for the treatment of hereditary angioedema. Expert Opinion on Orphan Drugs, 2015, 1, 141-156). For example, in the vitreous humor of patients with diabetic macular edema, overactivation of the KKS system has been found, leading to increased retinal vascular permeability and retinal thickening. In recent years, numerous studies have been published indicating that plasma kallikrein inhibitors can reduce retinal vascular permeability and are used to treat diabetic retinopathy and diabetic macular edema (Feener, EPlasma kallikrein and diabetic macular edema. Curr. Diab. Rep. 2010, 10, 270-275; Liu J. et al. Plasmakallikrein-kinin system and diabetic retinopathy. Biol. Chem. 2013, 394, 319-328).

[0004] Studies have confirmed that in healthy human volunteers, a dose-dependent decrease in plasma PKK levels is accompanied by a decrease in bradykinin production capacity, with acceptable safety and tolerability. This study reveals the potential for treating HAE and other diseases involving contact system activation and excessive bradykinin production by targeting PKK (Jason D.F. IONIS-PKKRx a Novel Antisense Inhibitor of Prekallikrein and Bradykinin Production. NUCLEIC ACID THERAPEUTICS, 2019, 29(2), Volume 29, 82-91).

[0005] Currently, no protein or peptide drugs targeting PPK have been successfully marketed. IONIS-PKK-LRx from IONIS Corporation, a 2￠-O-(2-methoxyethyl) modified chimeric antisense oligonucleotide, is designed to bind to and selectively reduce prekallikrein mRNA in the liver and is currently in Phase II clinical trials for the treatment of HAE. There are currently no antibodies or proteins that bind to plasma prekallikrein (PPK). Summary of the Invention

[0006] This invention provides an antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region includes the following three complementarity-determining regions (CDRs): CDR1 contains the sequence DYEMH; CDR2 contains the sequence AIHPGSGGTAYNQKFKG; and CDR3 contains the sequence PYGYPFAY; the light chain variable region includes the following three complementarity-determining regions (CDRs): CDR1 contains the sequence KASEDIYNRLA; CDR2 contains the sequence GETGLEA; and CDR3 contains the sequence QQYWSTPWT.

[0007] The antibody or antigen-binding fragments provided by this invention include murine antibodies, chimeric antibodies, humanized antibodies, or fully humanized antibodies or their antigen-binding fragments.

[0008] The present invention further provides some antibody or antigen-binding fragments, wherein the heavy chain variable region sequence is selected from any one of SEQ ID NO: 1-13, 15-23, and the light chain variable region sequence is selected from any one of SEQ ID NO: 24-42.

[0009] The present invention further provides antibody or antigen-binding fragments, wherein the heavy chain variable region is SEQ ID NO: 1, and the light chain variable region is selected from any one of SEQ ID NO: 24-32; or the heavy chain variable region is SEQ ID NO: 2 or 3, and the light chain variable region is selected from any one of SEQ ID NO: 24-26, 28 or 31; or the heavy chain variable region is SEQ ID NO: 4 or 6, and the light chain variable region is selected from any one of SEQ ID NO: 24-26, 28; or the heavy chain variable region is selected from any one of SEQ ID NO: 5, 7 or 8, and the light chain variable region is SEQ ID NO: 24; or the heavy chain variable region is selected from any one of SEQ ID NO: 9-13, and the light chain variable region is selected from any one of SEQ ID NO: 33-35; or the heavy chain variable region is SEQ ID NO: 15, and the light chain variable region is selected from any one of SEQ ID NO: 25, 36-42; or the heavy chain variable region is selected from SEQ ID NO: 15. NO: 12 or 16-23, the light chain variable region is SEQ ID NO: 42.

[0010] The antigen-binding fragments described in this invention are preferably derived from Fab, Fab'-SH, Fv, scFv, or (Fab')2 fragments.

[0011] The present invention also provides nucleic acid molecules encoding the aforementioned antibody or antigen-binding fragments; further provides expression vectors for expressing the nucleic acids; and further provides host cells containing the vectors, the host cells comprising prokaryotic or eukaryotic cells, more preferably selected from yeast cells, mammalian cells (e.g., 293 cells or CHO cells).

[0012] The nucleic acid molecules provided by this invention have heavy chain variable region nucleic acid sequences selected from any one of SEQ ID NO: 43-65, and light chain variable region nucleic acid sequences selected from any one of SEQ ID NO: 66-84.

[0013] The present invention further provides a nucleic acid molecule encoding a partial antibody or antigen-binding fragment, wherein the nucleic acid sequence of the heavy chain variable region is SEQ ID NO: 43, and the nucleic acid sequence of the light chain variable region is selected from any one of SEQ ID NO: 66-74; or the nucleic acid sequence of the heavy chain variable region is SEQ ID NO: 44 or 45, and the nucleic acid sequence of the light chain variable region is selected from any one of SEQ ID NO: 66-68, 70 or 73; or the nucleic acid sequence of the heavy chain variable region is SEQ ID NO: 46 or 48, and the nucleic acid sequence of the light chain variable region is selected from any one of SEQ ID NO: 66-68, 70; or the nucleic acid sequence of the heavy chain variable region is selected from any one of SEQ ID NO: 47, 49 or 50, and the nucleic acid sequence of the light chain variable region is SEQ ID NO: 66; or the nucleic acid sequence of the heavy chain variable region is selected from any one of SEQ ID NO: 51-55, and the nucleic acid sequence of the light chain variable region is selected from any one of SEQ ID NO: 75-77; or the nucleic acid sequence of the heavy chain variable region is selected from SEQ ID NO: 57, and the nucleic acid sequence of the light chain variable region is selected from SEQ ID NO: 66. The nucleic acid sequence of the heavy chain variable region is selected from any one of SEQ ID NO: 67, 78-84; or the nucleic acid sequence of the light chain variable region is selected from any one of SEQ ID NO: 54, 58-65, and the nucleic acid sequence of the light chain variable region is SEQ ID NO: 84.

[0014] The invention further provides light chain constant regions such as SEQ ID NO: 85 and its mutants; heavy chain constant regions such as Human IgG1, Human IgG2, Human IgG4, etc., wherein Human IgG1 is preferably SEQ ID NO: 86 and its mutants; Human IgG2 is preferably SEQ ID NO: 87 and its mutants; and Human IgG4 is preferably SEQ ID NO: 88 and its mutants.

[0015] The sequence correspondence between the antibody and antigen-binding fragments given in the examples of this invention is shown in Table 1:

[0016] Table 1. Sequences of some antibodies and antigen-binding fragments

[0017]

[0018]

[0019] The antibodies involved in the examples provided in this invention are all humanized antibodies, except for KH01, which is a murine antibody. The light chain constant region of the humanized antibody is SEQ ID NO: 85; the heavy chain constant region of KH02-KH30 and KH47-KH55 is Human IgG4 (SEQ ID NO: 88), and the heavy chain constant region of KH31-KH46 and KH56-KH63 is Human IgG1 (SEQ ID NO: 86).

[0020] In one specific embodiment, the antibody (KH34) involved in this invention has a heavy chain amino acid sequence as shown in SEQ ID NO: 89, a nucleotide sequence as shown in SEQ ID NO: 90, and a light chain amino acid sequence as shown in SEQ ID NO: 91, and a nucleotide sequence as shown in SEQ ID NO: 92.

[0021] The present invention also provides a pharmaceutical composition comprising the above-described antibody or antigen-binding fragment and a pharmaceutically acceptable excipient; the pharmaceutical composition is preferably an intravitreal injection formulation, a subretinal injection formulation, a choroidal injection formulation, an intravenous injection formulation, an intratumoral injection formulation, or an intramuscular injection formulation.

[0022] The present invention also provides the use of the aforementioned somatic or antigen-binding fragment or pharmaceutical composition in the preparation of a medicament for the prevention or treatment of diseases associated with plasma kallikrein or plasma prokallikrein in subjects; wherein the diseases associated with plasma kallikrein or plasma prokallikrein are preferably edema, rheumatoid arthritis, gout, intestinal diseases, stomatitis, neuropathic pain, inflammatory pain, spinal stenosis-degenerative spinal disease, diabetes, arterial or venous thrombosis, aortic aneurysm, osteoarthritis, vasculitis, pulmonary embolism, stroke, sepsis, systemic lupus erythematosus nephritis and burns, and retinal diseases; wherein the edema is preferably hereditary angioedema, cerebral edema, or head trauma; wherein the retinal diseases are preferably diabetic macular edema, retinal vein occlusion, age-related macular degeneration, macular edema secondary to retinal vein occlusion, uveitis, endophthalmitis, or polypoid choroidal vascular disease.

[0023] The MHL mentioned in this invention can be found in existing technologies (such as patents WO2011085103A, WO2014113701A, WO2017100679A, etc.) and can be constructed and prepared according to conventional technical means in the field. Its heavy chain is shown in sequence SEQ ID NO:14 and its light chain is shown in sequence SEQ ID NO:56.

[0024] definition

[0025] The term "binding" or "specific binding" as used in this invention refers to the binding of an antibody to an epitope of an antigen (such as human PPK) in an in vitro assay. The affinity of the binding is characterized by KD (binding rate) or Kd (dissociation constant). The antibody binding or specific binding of human plasma prokalopeptide-releasing enzyme (PPK) described in this invention does not preclude its possible binding to other antigens or epitopes, such as plasma kallikrein (PK).

[0026] In this invention, "epitope" refers to the portion of an antigen that specifically binds to an antibody. An epitope typically consists of chemically active (e.g., polar, nonpolar, or hydrophobic) surface groups of a moiety (such as an amino acid or polysaccharide side chain) and may possess specific three-dimensional structural features and specific charge characteristics. An epitope can be composed of continuous and / or discontinuous amino acids forming conformational spatial units. For discontinuous epitopes, amino acids from different portions of the linear sequence of the antigen are close together in three-dimensional space due to the folding of the protein molecule.

[0027] The term "antibody" as used in this invention refers to a binding protein containing an antigen-binding site. The term "binding site" or "antigen-binding site" refers to the region of the antibody molecule to which the ligand actually binds. The term "antigen-binding site" comprises an antibody heavy chain variable domain (VH) and / or an antibody light chain variable domain (VL) or a VH / VL pair, and may be derived from an intact antibody or antibody fragment such as a single-chain Fv, VH domain and / or VL domain, Fab, or (Fab)2. In one embodiment of the invention, each antigen-binding site comprises an antibody heavy chain variable domain (VH) and / or an antibody light chain variable domain (VL), and preferably consists of a pair of antibody light chain variable domains (VL) and antibody heavy chain variable domains (VH).

[0028] The antibody of the present invention may comprise six complementarity-determining regions (CDRs), including three heavy chain variable domain CDRs (CDRH1, CDRH2, and CDRH3) and three light chain variable domain CDRs (CDRL1, CDRL2, and CDRL3). CDRs and framework regions (FRs) together constitute the heavy or light chain variable domains. Amino acid residues in the “complementarity-determining regions” or “CDRs” are responsible for antigen binding. The “framework” or “FR” regions are the variable domain regions outside the complementarity-determining regions. FRs are less variable than CDRs; there are four FR molecules: FR1, FR2, FR3, and FR4. The light and heavy chains of the antibody, from the N-terminus to the C-terminus, include the domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The CDRs on each chain are separated by the framework amino acids. During antibody recognition, the four FR molecules coil to bring the CDR molecules closer together. In particular, CDR3 of the heavy chain is the region most conducive to antigen binding.

[0029] In this invention, the term "Fab" refers to a polypeptide comprising an antibody heavy chain variable domain (VH), an antibody constant domain 1 (CH1), an antibody light chain variable domain (VL), and an antibody light chain constant domain (CL), wherein the heavy and light chain domains are stabilized by disulfide bonds. One Fab can form one antigen-binding site, and two "Fab"s can form "(Fab')2" through disulfide bonds, thus having two antigen-binding sites. The "F(ab')2" fragment can be further reduced to form two Fab' fragments.

[0030] In this invention, the term "scFv" refers to a polypeptide containing an antibody heavy chain variable domain (VH) and a light chain variable domain (VL), which are stabilized by a short peptide. The scFv is a key region for antigen binding. "Fv" refers to a polypeptide containing an antibody heavy chain variable domain (VH) and a light chain variable domain (VL), which are linked together by non-covalent bonds.

[0031] The binding molecules or antibodies of the present invention further comprise immunoglobulin constant regions of one or more immunoglobulin species. Immunoglobulin species include IgG, IgM, IgA, IgD, and IgE isotypes, and in the case of IgG and IgA, include their subtypes. In a preferred embodiment, the antibody of the present invention has a constant domain structure of an IgG-type antibody.

[0032] The term "constant region" in this invention refers to the sum of antibody domains excluding variable regions. Constant regions are not directly involved in antigen binding but exhibit different effector functions. Antibodies are classified into the following categories based on the amino acid sequence of the constant region of the heavy chain: IgA, IgD, IgE, IgG, and IgM, with IgG and IgA further subdivided into the following subtypes: IgG1, IgG2, IgG3, and IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to different antibody classes are referred to as α, δ, ε, γ, and μ, respectively. The light chain constant regions of all five antibody classes are referred to as κ (kappa) and λ (lambda). In this invention, constant regions derived from humans refer to the heavy chain constant regions and / or κ or λ light chain constant regions of human antibodies subclasses IgG1, IgG2, IgG3, or IgG4.

[0033] The antibody or antigen-binding fragments according to the present invention also include binding molecules or antibodies ("variants") having "conserved sequence modifications." This means nucleotide and amino acid sequence modifications that do not affect or alter the aforementioned characteristics. Nucleotides or amino acids can be modified using techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Non-essential amino acid residues in the antibody or antigen-binding fragment can preferably be substituted with another amino acid residue from the same side chain family. Therefore, a "variant" antibody or antigen-binding fragment refers to an amino acid sequence having one or more amino acid additions, deletions, and / or substitutions compared to the "parent" amino acid sequence. Therefore, the nucleotide or amino acid sequences of the antibody or antigen-binding fragments of the present invention all include sequences having "conserved sequence modifications" with at least 80%, 85%, 90%, 95%, 98%, 99%, or higher homology to the parent nucleotide or amino acid.

[0034] The term "chimeric antibody" in this invention refers to an antibody comprising an antibody variable domain from one source or species and at least a portion of a constant region derived from another source or species, typically prepared using recombinant DNA technology. For example, in one embodiment, a chimeric antibody comprising a mouse variable region and a human constant region.

[0035] In this invention, the term "humanized antibody" refers to an antibody whose constant region (i.e., the CH and CL regions) or which is entirely encoded by a human antibody gene. Humanized antibodies can significantly reduce the immune response induced by heterologous antibodies in the human body.

[0036] The term "host cell" in this invention refers to any cell line that can be modified to produce the antibodies described in this invention. In one embodiment, HEK293 cells and CHO cells are used as host cells. In this application, the terms "cell," "cell line," and "cell culture" are used interchangeably and include their progeny.

[0037] The term "pharmaceutically acceptable excipient" in this invention refers to any formulation or carrier medium capable of delivering an effective amount of the active molecule of this invention without interfering with the biological activity of the active molecule and without toxic side effects on the host or patient. This includes any solvents, dispersion media, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonics, absorption delay agents, salts, preservatives, pharmaceuticals, pharmaceutical stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, and combinations thereof known to those skilled in the art. Preferably, the excipient is suitable for administration via intraocular, intravenous, intramuscular, subcutaneous, parenteral, or intra-articular routes.

[0038] Instruction manual illustrations

[0039] Figure 1Bands from partial SDS-PAGE for PK precursor activity detection

[0040] Figure 2 Scoring criteria for fundus fluorescein angiography in rhesus monkeys (scores range from 0 to 10 based on the severity of retinal microvascular leakage; 0 for the saline group, 8 for the group given only CA-I, 9 for incomplete light transmission and blurred fundus, and 10 for complete lack of light transmission (animal has no light perception)).

[0041] Figure 3 Scoring results for the severity of microvascular leakage in the retina of rhesus monkeys

[0042] Figure 4 Fundus fluorescein angiography image of rhesus monkeys Detailed Implementation

[0043] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0044] The antibody heavy and light chain CDR transplantation, PCR introduction of site mutations, and screening of mutant libraries described in this invention are accomplished through conventional gene recombination technology and immunological techniques based on antigen-antibody interactions. Specific experimental methods and steps are as described in <<Molecular Cloning>> 3rd Edition (Joseph Sambrook, Science Press, August 1, 2002) and similar experimental manuals.

[0045] Example 1: Mouse Immunization and Hybridoma Screening: BALB / C female mice (6-8 weeks old, purchased from Dashuo Laboratory Animal Co., Ltd.) were immunized using either standard intraperitoneal or plantar immunization methods. Each mouse received 10 μg of Human kallikrein antigen (Enzyme Research, catalog number: 3792A) every 2-3 weeks. For the initial immunization, all mice were immunized with an equal volume of CFA (SIGMA, catalog number: SLBW7430) and Human kallikrein antigen (Enzyme Research, catalog number: 3792A), emulsified together. Booster immunizations before the sprint immunization (approximately the second to fourth immunizations) were performed using IFA (SIGMA, catalog number: F5506). Three days before fusion, mice underwent a sprint immunization. The Human kallikrein antigen used for the pre-fusion sprint immunization was prepared with PBS (20 μg / mouse). Mice were euthanized by twisting their necks and disinfected by immersion in 70% ethanol for 5 minutes. Spleen and lymph nodes were harvested, and the spleen and lymph nodes were appropriately ground. The obtained B lymphocytes or spleen B cells were then mixed with P3X63Ag8.653 myeloma cells (product number: CRL-1580 (purchased from ATCC) or FO myeloma cells (catalog number: TCM31, purchased from the Chinese Academy of Sciences Cell Bank) were mixed at a certain ratio and fused using an electrofusion instrument. When the hybridoma cells recovered to their optimal state, ELISA was performed. Based on the ELISA results, positive hybridoma cells were selected for expanded culture. A second test (combining ELISA and functional activity) was performed 2-3 days later, and functionally active hybridoma cell lines were expanded. Total RNA was extracted from hybridoma cells using the Trizol method, and 5 μg of total RNA was reverse transcribed to obtain cDNA. Then, using cDNA as a template, PCR amplification was performed using 5' RACE technology. The PCR product was ligated into the restriction enzyme vector pcDNA3.4, and the ligation product was transformed into competent Stellar Competent Cells by heat shock. Colony PCR identification was performed, and a small amount of plasmid was extracted from positive clones for sequencing to obtain the light and heavy chain variable region sequences of the hybridoma antibody gene.

[0046] Example 2: Construction of Chimeric Antibody: Primers were designed based on the variable region gene sequence of mouse hybridoma antibody and the constant region of human light and heavy chains. The variable region gene sequence of mouse hybridoma antibody was amplified by PCR to obtain the chimeric antibody Q fragment. The constant regions of human light and heavy chains (hIgG4 or hIgG1) were amplified by PCR to obtain the chimeric antibody H fragment. The Q and H fragments were simultaneously ligated into the restriction enzyme vector pcDNA3.4 using homologous recombination. The correctly sequenced plasmid was transfected into expiCHO or HEK293 cells for protein expression. Cells were cultured for approximately 7-10 days, and the cell supernatant was collected by centrifugation. The cell supernatant was purified using an AKTA protein purifier to obtain the chimeric antibody protein.

[0047] Example 3: Antibody Humanization: Since murine antibodies easily induce HAMA reactions in humans, humanization is necessary to reduce their immunogenicity in humans. This example employs traditional methods such as CDR transplantation, framework reorganization, and key amino acid reverse mutation design to humanize multiple candidate molecules. Specifically, the method involves first comparing the CDR regions of the murine antibody light and heavy chains with the FR region of a highly similar human germline gene, obtaining the humanized antibody variable regions. Then, homologous recombination is used to splice the constant regions of the light and heavy chains with the corresponding antibody variable regions and ligate them into expression vectors. Colony PCR screening and sequencing are then used to identify the correct antibody light and heavy chain expression plasmids. The correctly sequenced light and heavy chain plasmids are then transfected into expiCHO or HEK293 cells for expression. After 7-10 days, the cell supernatant is purified using an AKTA protein purifier to obtain the humanized antibody protein. The humanized antibody activity is then measured. The selected antibody molecules were compared with the parent antibody molecules in sequence. Based on factors such as amino acid position, polarity, molecular weight, and spatial conformation, the amino acid molecules that have a greater impact on activity were identified. These amino acids were then reverse-mutated to further increase the activity of the antibody molecules. Humanized antibody molecules were finally obtained through combination screening.

[0048] Example 4: Binding Affinity Test with Human PPK

[0049] The affinity of the candidate antibody for the antigen Human prekallikrein (Enzymeresearch, catalog number: HPK1302) was detected using the Octet QKe refurbisher. The candidate antibody was loaded onto the sensor and subjected to binding and dissociation reactions with different concentrations of antigen under the following conditions: baseline time: 120 s, loading time: 300 s, association time: 600 s, and dissociation time: 2000 s. After the reaction was completed, the results were analyzed using Octet analysis software, and the results are shown in Table 2.

[0050] Table 2

[0051]

[0052] Example 5: Detection Method for Inhibiting the Conversion of PPK to PK

[0053] Plasma pre-Kallikrein (PPK) is a single-amino acid protein (molecular weight 86 kDa). When activated by Factor XIIa cleavage (cleaving only one peptide bond), it becomes enzymatically active PlasmaKallikrein (PK), still with a molecular weight of 86 kDa, but consisting of a single disulfide-linked double peptide chain (52 kDa + 36 / 33 kDa). Reduction SDS-PAGE clearly distinguishes PPK and PK proteins, allowing for the detection of whether antibody molecules can inhibit Factor XIIa cleavage of PPK into PK. The sample reaction system is 15 μL, mainly containing 250 μg / mL PPK protein, 250 ng / mL Factor XIIa, and 500 μg / mL antibody protein. The diluent is the detection buffer. First, prepare the PPK protein and antibody protein samples with detection buffer and incubate at 37°C for 45 min. Then, add Factor XIIa and incubate at 37°C for another 45 min. After the reaction, add 5 μl of 4× Loading buffer and incubate at 100°C for 10 min to terminate the reaction. Next, perform SDS-PAGE electrophoresis, loading 10 μl per well onto a 5% stacking gel (80V, 40 min) and a 12% separating gel (120V, 1 h), allowing the bromophenol blue to reach the bottom of the gel. After electrophoresis, remove the gel, rinse with purified water, and stain with SDS-PAGE instant blue staining solution for 5-15 min. After staining, destain with purified water until the background color lightens. Finally, take images using an ImageLab gel imaging system.

[0054] Figure 1 The results showed that PPK is a single-amino acid protein with a molecular weight of 86 kDa, and reduced SDS-PAGE showed a single band (see lane 1). Upon addition of F12a factor, PPK was cleaved into PK, which also had a molecular weight of 86 kDa and consisted of two disulfide-linked peptide chains. After protein denaturation, PK separated into peptide chains with molecular weights of 52 kDa and 36 / 33 kDa, respectively (see lane 2). The addition of antibody molecules significantly inhibited F12a factor cleavage of PPK (see lanes 3-8); however, MHL, which lacks PPK activity, could not inhibit PPK cleavage (see lane 9). KHa-e was a murine antibody molecule obtained through hybridoma screening as described in Example 1.

[0055] Example 6: Method for Detecting Human Plasma Activity

[0056] Human plasma contains proteins such as PPK and PK, but detection has shown that PK does not exhibit activity in either undiluted or diluted human plasma, possibly indicating an equilibrium state of enzyme inhibition. Adding Factor XIIa disrupts this equilibrium, producing PK protein with enzymatic activity. This principle is used to detect the activity of PK inhibitors. The first step is antibody sample preparation. The antibody protein is diluted to 200 μg / ml using detection buffer (20 mM Tris-HCl, pH 7.50, 150 mM NaCl, 1 mM EDTA, 0.1% PEG-8000, and 0.1% Triton X-100) as the starting concentration. A four-fold serial dilution is then performed, resulting in 11 concentration gradients. The diluted human plasma is then diluted 40-fold using detection buffer, Factor XIIa is diluted to 100 ng / ml, and the substrate peptide (H-Pro-Phe-Arg-AMC) is diluted to 1000 μM for later use. Add 50 μl of detection buffer and diluted antibody protein solutions of various concentrations to each well of a 96-well opaque ELISA plate; add 20 μl of 40-fold diluted human plasma to each well of the same 96-well plate and incubate at 37°C for 30 min; then add 20 μl of 100 ng / ml Factor XIIa and incubate at 37°C for 45 min; finally, add 10 μl of 1000 μM substrate peptide (H-Pro-Phe-Arg-AMC) to each well of the 96-well opaque ELISA plate. Place the prepared 96-well plate into a multi-functional microplate reader (Spectra Max i3X) for detection. Detection parameters: excitation light 360 nm, emission light 480 nm, read every 60 s for a total of 10 min. Normalize the enzyme reaction rate (slope) and perform a four-parameter curve fitting with the concentration (nM) to calculate the IC50 (nM). Specific results are shown in Table 3.

[0057] Table 3

[0058]

[0059]

[0060] Example 7: Animal Efficacy Study

[0061] 1. Construction of primate disease models

[0062] In the vitreous body of patients with diabetic retinopathy, there is significant overexpression of carbonic anhydrase I (CA-I). Increased CA-I expression alters the vitreous microenvironment, thereby activating the KKS signaling pathway, leading to increased retinal vascular permeability, resulting in retinal vascular leakage and macular edema (DME). In rhesus monkeys (4 years old, 3.2-4.9 kg), intravitreal injection of carbonic anhydrase at doses of 0.4 mg–0.55 mg / eye was performed, and fluorescein fundus angiography (FFA) 30 minutes later revealed more severe microvascular leakage.

[0063] 2. The excipient formulations containing the test drug are shown in Table 3:

[0064] Table 3 Excipient formulations of the test drugs

[0065] sucrose 50mg / ml Arginine 17.42 mg / ml Twain 20 0.5mg / ml Citric acid 2.1mg / ml hydrochloric acid Adjust the pH to 7.5.

[0066] 3. Administration method

[0067] The test drug (KH34) was administered intravitreally at doses of 25 μg / eye, 5 μg / eye, and 1 μg / eye. The following day, intravitreal injection of carbonic anhydrase was performed to establish the retinal vascular leakage model. Thirty minutes after model establishment, fundus fluorescein angiography (FFA) was used to observe the inhibitory effect of the drug molecule on CA-I-induced retinal vascular leakage, evaluating the efficacy of the candidate molecule in a primate DME disease model.

[0068] 4. Experimental Results

[0069] Scoring criteria: The severity of retinal microvascular leakage was scored from 0 to 10. The saline group received 0 points for FFA images, the CA-I-only group received 8 points, incomplete light transmission and blurred fundus were 9 points, and complete lack of light transmission (animal had no light perception) was 10 points. See specific images. Figure 2 .

[0070] The experimental study scoring results are shown below. Figure 3 Fundus fluorescein angiography showed Figure 4 (4 eyes per group of test drugs) The drug of this invention can significantly inhibit retinal vascular leakage. sequence list <110> Chengdu Kanghong Biotechnology Co., Ltd. <120> An antibody and its uses <130> KH20220106 <150> 2021101018271 <151> 2021-01-25 <160> 92 <170> SIPOSequenceListing 1.0 <210> 1 <211> 117 <212> PRT <213> Artificial sequence <400> 1 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Leu Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Lys Gln Thr Pro Val Leu Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Ile Phe Thr Ala Asp Lys Ser Ser Arg Thr Ala Tyr 65 70 75 80 Met Tyr Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ala 115 <210> 2 <211> 117 <212> PRT <213> Artificial sequence <400> 2 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 3 <211> 117 <212> PRT <213> artificial sequence <400> 3 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 4 <211> 117 <212> PRT <213> Artificial sequence <400> 4 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 5 <211> 117 <212> PRT <213> artificial sequence <400> 5 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 6 <211> 117 <212> PRT <213> artificial sequence<o000285><400> 6 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala[[ID=SO]] 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 7 <211> 117 <212> PRT <213> artificial sequence <400> 7 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala[[ID=二十八]] 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 8 <211> 117 <212> PRT <213> artificial sequence[[ID=佃]] <400> 8 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe[[ID=孔]] 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 9 <211> 117 <212> PRT <213> artificial sequence <400> 9 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Leu Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 10 <211> 117 <212> PRT <213> Artificial sequence <400> 10 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 11 <211> 117 <212> PRT <213> Artificial sequence <400> 11 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60<000​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Arg Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 13 <211> 117 <212> PRT <213> artificial sequence <400> 13 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 14 <211> 451 <212> PRT <213> MHL H <400> 14 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 His Tyr 20 25 30 Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Tyr Ser Ser Gly Gly Ile Thr Val 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 Tyr Arg Arg Ile Gly Val Pro Arg Arg Asp Glu 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 Arg Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 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 450 <210> 15 <211> 117 <212> PRT <213> artificial sequence <400> 15 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 16 <211> 117 <212> PRT <213> artificial sequence <400> 16 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 17 <211> 117 <212> PRT <213> artificial sequence <400> 17 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Met Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 18 <211> 117 <212> PRT <213> artificial sequence <400> 18 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 19 <211> 117 <212> PRT <213> artificial sequence <400> 19 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Val Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 20 <211> 117 <212> PRT <213> artificial sequence <400> 20 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Leu Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 21 <211> 117 <212> PRT <213> artificial sequence <400> 21 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Ile Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 22 <211> 117 <212> PRT <213> artificial sequence <400> 22 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Thr Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 23 <211> 117 <212> PRT <213> artificial sequence <400> 23 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met[[ID=I]] 35 40 45 Gly Ala Ile His Pro Gly Ser Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Tyr Gly Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 24 <211> 107 <212> PRT <213> artificial sequence <400> 24 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Phe Ser Gly Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Met 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Leu Ser Ile Ala Ser Leu Gln Thr 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 25 <211> 107 <212> PRT <213> artificial sequence <400> 25 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 Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 26 <211> 107 <212> PRT <213> artificial sequence <400> 26 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 Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 27 <211> 107 <212> PRT <213> Artificial sequence <400> 27 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 Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 28 <211> 109 <212> PRT <213> Artificial sequence <400> 28 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 <210> 29 <211> 107 <212> PRT <213> Artificial sequence <400> 29 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly[[ID=�5]] 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 30 <211> 107 <212> PRT <213> artificial sequence <400> 30 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 31 <211> 107 <212> PRT <213> artificial sequence <400> 31 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 32 <211> 107 <212> PRT <213> Artificial sequence <400> 32 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 33 <211> 107 <212> PRT <213> Artificial sequence <400> 33 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 Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 34 <211> 107 <212> PRT <213> artificial sequence <400> 34 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 35 <211> 107 <212> PRT <213> artificial sequence <400> 35 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 Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Met 35 40 45 Ser Gly Glu Thr Gly Leu Glu Ala 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 Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 36 <211> 107 <212> PRT <213> artificial sequence <400> 36 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Leu 35 40 45 Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys His Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 37 <211> 107 <212> PRT <213> Artificial sequence <400> 37 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Leu 35 40 45 Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly [[ID=K]]50 55 60 Ser Gly Ser Gly Thr His Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 38 <211> 107 <212> PRT <213> Artificial sequence <400> 38 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Leu 35 40 45 Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr His Tyr Thr Leu Thr Ile Ser Ser Leu Gln Thr 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 39 <211> 107 <212> PRT <213> artificial sequence <400> 39 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg ​​​​​​Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr His Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 40 <211> 107 <212> PRT <213> artificial sequence <400> 40 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Leu 35 40 45 Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95[[ID=...]] / / 此处省略号表示未翻译的标签,下同 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 41 <211> 107 <212> PRT <213> artificial sequence <400> 41 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Leu 35 40 45 Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr His Tyr Thr Leu Thr Ile Ala Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 42 <211> 107 <212> PRT <213> Artificial sequence <400> 42 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Leu 35 40 45 Tyr Gly Glu Thr Gly Leu Glu Ala Gly Phe His Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr His Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys<00?1061>100 105 <210> 43 <211> 351 <212> DNA <213> Artificial sequence <400> 43 caggttcaac tgcagcagtc tggggctgaa ctgatgaggc ctggggcttc agtgaaggtg 60 tcctgcaagg ctttgggcta cacatttact gactatgaaa tgcactgggt gaaacagaca 120 cctgtgcttg gcctggaatg gattggagct attcatccag gaagtggtgg tactgcctac 180 aatcagaagt tcaagggcaa ggccatattc actgcagaca aatcctccag gacagcctac 240 atgtacctca gcagcctgac atctgaggac tctgctgtct attactgtac aagaccgtat 300 ggttaccctt ttgcttactg gggccaaggg actctggtca ctgtctctgc a 351 <210> 44 <211> 351 <212> DNA <213> Artificial Sequence <400> 44 caagtgcaac tagtgcaaag tggtagtgaa ctgaagaagc ctggcgcttc cgtgaaggtg 60 tcctgcaagg cctctggata caccttcacc gactacgaga tgcactgggt gcggcagcct 120[[ID=2,3]] cctggaaaag gcctggagtg gatcggcgcc atccaccccg gctctggcgg cacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgccgaca agtccacctc taccgcttac 240 atggaactgt ccagcctgcg gtccgaggat acagccgtgt actactgtgc tagaccttac 300 ggctatccat ttgcctactg gggccagggc accctggtca ccgtgtctag c 351 <210> 45 <211> 351 <212> DNA <213> Artificial sequence <400> 45 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggcaaag gcctggaatg gatcggcgcc atccaccccg gatccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccaccag caccgcctac 240 atggaactgt ccagcctgcg gtccgaggac accgccgtgt actactgtgc tagaccttac 300 ggctatcctt ttgcctactg gggccagggc accctggtga cagtgtcttc t 351 <210> 46 <211> 351 <212> DNA <213> Artificial sequence <400> 46 caagtgcaac tagtgcaaag tggtagtgaa ctgaagaagc ctggcgctag cgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcaggct 120 cctggaaaag gcctggagtg ggtcggcgcc atccaccccg gctccggcgg aacagcctac 180 aaccagaagt tcaagggcag agtgaccatc accgccgaca agtccacctc tacagcttac 240 atggaactgt cctctctgcg gtccgaggat accgctgtgt actactgtgc cagaccttac 300 ggctatccat ttgcctactg gggccagggc accctggtga ccgtgtctag c 351 <210> 47 <211> 351 <212> DNA <213> Artificial sequence <400> 47 caagtgcaac tagtgcaaag tggtgcagaa gtgaagaagc ctggcgcttc cgtgaaagtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcaggct 120 cctggcaagg gcctggagtg ggtcggcgcc atccaccccg gctccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgccgaca agtccacctc tacagcctac 240 atggaactga gctctctgcg gtccgaggat accgctgtgt actactgtgc cagaccttac 300 ggctaccctt ttgcctactg gggccagggc accctggtga ccgtgtctag c 351 <210> 48 <211> 351 <212> DNA <213> Artificial sequence <400> 48 caagtgcaac tagtgcaaag tggtgcagaa gtgaagaagc ctggcgcttc cgtgaaagtg 60 caagtgcaac tagtgcaaag tggtgcagaa gtgaagaagc ctggcgcttc cgtgaaagtg 60 tcctgcaagg cctccggcta caccttcacc gactacgaga tgcactgggt gcggcaggct 120 tcctgcaagg cctccggcta caccttcacc gactacgaga tgcactgggt gcggcaggct 120 cctggcaagg gcctggaatg ggtcggcgcc atccaccccg gatctggcgg aacagcttac 180 cctggcaagg gcctggaatg ggtcggcgcc atccaccccg gatctggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccacctc tacagcctac 240 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccacctc tacagcctac 240 atggaactga gctctctgcg gtccgaggac accgccgtgt actactgtgc cagaccttac 300 atggaactga gctctctgcg gtccgaggac accgccgtgt actactgtgc cagaccttac 300 ggctatccat ttgcctactg gggccagggc accctggtga ccgtgtctag c 351 ggctatccat ttgcctactg gggccagggc accctggtga ccgtgtctag c 351 <210> 49<210> 49 <211> 351<211> 351 <212> DNA<212> DNA <213> 人工序列(artifical sequence) <213> Artificial sequence <400> 49 <400> 49 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctccggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 tcctgcaagg cctccggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggcaaag gcctggagtg gatcggcgcc atccaccctg gatctggcgg aacagcttac 180 ccaggcaaag gcctggagtg gatcggcgcc atccaccctg gatctggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgccgaca agtccacctc taccgcttac 240 aaccagaagt tcaagggcag agtgaccatc accgccgaca agtccacctc taccgcttac 240 atggaactgt ccagcctgcg gtccgaggat acagccgtgt actactgtgc tagaccttac 300 ggctatccct ttgcctactg gggccagggc accctggtga ccgtgtctag c 351 <210> 50 <211> 351 <212> DNA <213> Artificial sequence <400> 50 caagtgcaac tagtgcaaag tggtagtgaa ctgaagaagc ctggagcttc cgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 cctggcaaag gcctggaatg gatcggcgcc atccaccccg gctccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agagcacatc taccgcctac 240 atggaactgt ccagcctgcg gtccgaggac accgccgtgt actactgtgc tagaccttac 300 ggctatccat ttgcctactg gggccagggc accctggtca ccgtgtcttc t 351 <210> 51 <211> 351 <212> DNA <213> Artificial sequence <400> 51 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggccttg gcctggaatg gatcggcgcc atccaccccg gatccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccaccag caccgcctac 240 atggaactgt ccagcctgcg gtccgaggac accgccgtgt actactgtgc tagaccttac 300 ggctatcctt ttgcctactg gggccagggc accctggtga cagtgtcttc t 351 <210> 52 <211> 351 <212> DNA <213> Artificial Sequence <400> 52 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggcaaag gcctggaatg gatcggcgcc atccaccccg gatccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccaccag caccgcctac 240 atggaactgt ccagcctgac atccgaggac accgccgtgt actactgtgc tagaccttac 300 ggctatcctt ttgcctactg gggccagggc accctggtga cagtgtcttc t 351 <210> 53 <211> 351 <212> DNA <213> artificial sequence <400> 53 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggcaaag gcctggaatg gatcggcgcc atccaccccg gatccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccaccag caccgcctac 240 atggaactgt ccagcctgcg gtccgaggac tctgccgtgt actactgtgc tagaccttac 300 ggctatcctt ttgcctactg gggccagggc accctggtga cagtgtcttc t 351 <210> 54 <211> 351 <212> DNA <213> artificial sequence <400> 54 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggcaaag gcctggaatg gatcggcgcc atccaccccg gatccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgatg agtccaccag gaccgcctac 240 atggaactgt ccagcctgcg gtccgaggac accgccgtgt actactgtgc tagaccttac 300 ggctatcctt ttgcctactg gggccagggc accctggtga cagtgtcttc t 351 <210> 55 <211> 351 <212> DNA <213> Artificial sequence <400> 55 caagtgcaac tagtgcaaag tggtgcagaa gtcaagaagc ctggcgcttc tgtgaaggtg 60 tcctgcaagg cctctggcta caccttcacc gactacgaga tgcactgggt gcggcagcct 120 ccaggcaaag gcctggaatg gatcggcgcc atccaccccg gatccggcgg aacagcttac 180 aaccagaagt tcaagggcag agtgaccatc accgctgata aatccaccag caccgcctac 240 atggaactgt ccagcctgcg gtccgaggac accgccgtgt actactgtgc tagaccttac 300 ggctatcctt ttgcctactg gggccagggc accctggtga cagtgtcttc t 351 <210> 56 <211> 213 <212> PRT <213> MHL <400> 56 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 Thr 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 Asn Thr Tyr Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile 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> 57 <211> 351 <212> DNA <213> artificial sequence <400> 57 caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 60 tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120 cctggacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 58 <211> 351 <212> DNA <213> Artificial Sequence <400> 58 caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggcgcctc tgtgaaggtg 60 tcttgtaagg cttctggata cacattcaca gattatgaaa tgcactgggt gagacaggcc 120 cctggacaga gactggaatg gatgggcgct atccatcctg gatctggcgg aacagcctac 180 aaccagaagt ttaagggaag agtgacaatt acaagagata caagcgcttc tacagcttac 240 atggagctga gctctctgag atctgaggat acagctgtgt actactgtgc tagaccttac 300 ggataccctt ttgcttactg gggccagggc acactggtga ccgtgagctc t 351 <210> 59 <211> 351 <212> DNA <213> Artificial Sequence <400> 59 caggtgcagc tggtgcagtc tggcgctgag gtgatgaagc ctggagcctc tgtgaaggtg 60 tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120 cctggacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 60 <211> 351 <212> DNA <213> Artificial sequence <400> 60 caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 60 tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120 cctggacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgac atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 61 <211> 351 <212> DNA <213> Artificial sequence <400> 61 caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 60caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 60 tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120 cctgtacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180cctgtacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 62<210> 62 <211> 351<211> 351<00012atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 63 <211> 351 <212> DNA <213> Artificial sequence <400> 63 caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 6o tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120 cctggacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgataatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 64 <211> 351 <212> DNA <213> Artificial sequence <400> 64 caggtgcagc tggtgcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 60 It should be noted that there is a small error in the original text where "6o" in line should probably be "60". This has been corrected in the translation.tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacagact 120 cctggacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 65 <211> 351 <212> DNA <213> Artificial sequence <400> 65 caggtgcagc tgcagcagtc tggcgctgag gtgaagaagc ctggagcctc tgtgaaggtg 60 tcttgtaagg cctctggcta cacattcaca gattatgaga tgcactgggt gagacaggct 120 cctggacagg gcctggagtg gatgggagct atccaccctg gctctggagg aacagcttac 180 aaccagaagt ttaagggaag agtgacaatg acaagagata catctacatc tacagtgtat 240 atggagctga gctctctgag atctgaggat accgccgtgt actattgtgc tagaccttat 300 ggctatcctt ttgcttactg gggacagggc acactggtga ccgtgtctag c 351 <210> 66 <211> 321 <212> DNA <213> Artificial sequence <400> 66 gacatccaga tgacacaatc ttcatcctcc ttttctggat ctctaggaga cagagtcacc 60 attacttgca aggcaagtga ggacatatat aatcggttag cctggtatca gcagaaacca 120 ggaaatgctc ctaggctctt aatgtctggt gagaccggtt tagaagctgg gtttcattta 180 agattcagtg gcagtggatc tggaaaggat tacactctca gcattgccag tcttcagact 240 gaagatgttg ctacttatta ctgtcaacag tattggagta ctccgtggac gttcggtgga 300 ggcaccaagc tggaaatcaa a 321 <210> 67 <211> 321 <212> DNA <213> Artificial sequence <400> 67 gatattcaaa tgactcaaag tccaagtagt ctgtctgcct ccgtgggcga cagagtgacc 60 atcacctgta aggcctctga ggacatctac aaccggctgg cctggtacca gcagaagccc 120 ggacaggctc ctagactgct gatcagcggc gagacaggcc tggaagctgg cgtcccatct 180 cggttctccg gctccggctc tggcaccgac ttcaccctga ccatctccag cctccagcct 240 gaggatgtgg ccacctacta ctgccagcag tattggtcca ccccttggac ctttggccaa 300 ggaacaaaag tggaaatcaa g 321 <210> 68 <211> 321 <212> DNA[[ID=Ill]] <213> Artificial sequence <400> 68 gatattcaaa tgactcaaag tccaagtagt ctctctgctt ccgtgggcga cagagtgacc 60 atcacctgca aggcctccga ggacatctac aaccggctgg cctggtatca gcagaaacca 120 ggccaggccc ctagactgct gatctctggc gagacaggcc tggaagctgg cgtccccagc 180 cggttctccg gctctggatc tggaacagac ttcaccctga ccatctccag cctgcagcct 240 gaggattttg ccacctacta ctgtcaacag tactggtcca ccccttggac cttcggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 69 <211> 321 <212> DNA <213> Artificial sequence <400> 69 gatattcaaa tgactcaaag tccaagtagt ctgtctgcct ctgtcggcga cagagtgaca 60 atcacctgta aagcctccga ggacatctac aaccggctgg cctggtatca gcaaaagcct 120 ggcaaggccc caaagctgct gatcagcggc gagacaggcc tggaagctgg cgtgccctct 180 agattctccg gatctggctc cggaaccgac ttcaccctga ccatctccag cctccagcct 240 gaggattttg ctacctacta ctgccagcag tactggtcca ccccttggac cttcggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 70 <211> 321 <212> DNA <213> Artificial Sequence <400> 70 gatattcaaa tgactcaaag tccaagtagt gtcagcgcct ccgtgggcga cagagtgacc 60 atcacctgta aggcctctga ggacatctac aaccggctgg cctggtatca gcagaaacca 120 ggccaagctc ctagactgct gatctctggc gagacaggac tggaagctgg cgtgcccagc 180 cggttctccg gctccggctc tggaaccgac tttacactga ccatctcctc tctgcagcct 240 gaggatttcg ccacctacta ctgccagcag tactggtcca ccccttggac cttcggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 71 <211> 321 <212> DNA <213> Artificial sequence <400> 71 gatattcaaa tgactcaaag tccaagtagt ctgtccgcct ccgtgggcga cagagtgacc 60 atcacctgca aggcctctga ggacatctac aaccggctgg cctggtatca gcagaagcct 120 ggcaaagctc caaagctcct gatcagcggc gagacaggcc tggaagctgg cgtccccagc 180 agattctccg gctctggatc tggaaccgac ttcaccctga ccatctcctc tctgcagcct 240 gaggatgtgg ccacctacta ctgtcaacag tactggtcca caccttggac ctttggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 72 <211> 321[[ID=2零]] <212> DNA <213> Artificial sequence <400> 72 gatattcaaa tgactcaaag tccaagtagt gtgtccgcct ccgtgggcga cagagtgaca 60 atcacctgta aagcctctga ggacatctac aaccggctgg cctggtatca gcagaagcct 120 ggccaagctc caagactgct gatctctggc gagaccggcc tggaagctgg agtccccagc 180 cggttttccg gctctggatc tggcaccgac ttcaccctga caatctccag cctgcagcct 240 gaggatgtgg ccacctacta ctgccagcag tactggtcca ccccttggac cttcggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 73 <211> 321 <212> DNA <213> Artificial sequence <400> 73 gatattcaaa tgactcaaag tccaagtagt gtcagcgcct ctgtgggcga cagagtgacc 60 atcacctgca aggcctccga ggacatctac aaccggctgg cctggtatca gcagaagcct 120 ggcaaagctc caaagctgct gatctctggc gagacaggcc tggaagctgg cgtgccctcc 180 agattctccg gctctggatc tggaacagac ttcaccctga ccatctccag cctgcagcct 240 gaggatgtgg ccacctacta ctgtcaacag tactggtcca ccccttggac ctttggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 74 <211> 321 <212> DNA <213> Artificial sequence <400> 74 gatattcaaa tgactcaaag tccaagtagt gtcagcgcct ctgtgggcga cagagtgaca 60 atcacctgca aggcctccga ggacatctac aaccggctgg cctggtatca gcagaagcct 120 ggcaaagctc caaagctgct gatctctggc gagacaggcc tggaagctgg cgtgccctcc 180 agattctccg gctctggatc tggaaccgac ttcaccctga ccatctccag cctgcagcct 240 gaggattttg ccacctacta ctgtcaacag tactggtcca ccccttggac cttcggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 75 <211> 321 <212> DNA <213> Artificial sequence <400> 75 gatattcaaa tgactcaaag tccaagtagt ctctctgctt ccgtgggcga cagagtgacc 60 atcacctgca aggcctccga ggacatctac aaccggctgg cctggtatca gcagaaacca 120 ggccaggccc ctagactgct gatctctggc gagacaggcc tggaagctgg cgtccccagc 180 cggttctccg gctctggatc tggaacagac tataccctga ccatctccag cctgcagcct 240 gaggattttg ccacctacta ctgtcaacag tactggtcca ccccttggac cttcggccag 300 ggcaccaagg tggaaatcaa g 321 <210> 76 <211> 321 <212> DNA <213> Artificial sequence <400> 76 gatattcaaa tgactcaaag tagtagtagt ctctctgctt ccgtgggcga cagagtgacc atcacctgca aggcctccga ggacatctac aaccggctgg cctggtatca gcagaaacca 180. ggccaggccc ctagactgct gatctctggc gagacaggcc tggaagctgg cgtccccagc cggttctccg gctctggatc tggaacagac ttcaccctga ccatctccag cctgcagcct 240 gaggattttg ccacctacta ctgtcaacag tactggtcca ccccttggac cttcggccag ggcaccaagg tggaatcaa g <210> 77 <211> 321 <212> DNA <213> Artificial sequence (artificial sequence) <400> 77 gatattcaaa tgactcaaag tccaagtagt ctctctgctt ccgtgggcga cagagtgacc atcacctgca aggcctccga ggacatctac aaccggctgg cctggtatca gcagaaacca ggccaggccc ctagactgct gatgtctggc gagacaggcc tggaagctgg cgtccccagc 180 cggttctccg gctctggatc tggaacagac ttcaccctga ccatctccag cctgcagcct 240 gaggattttg ccacctacta ctgtcaacag tactggtcca ccccttggac cttcggccag ggcaccaagg tggaaatcaa g 321 <210> 78 <211> 321 <212> DNA <213> Artificial sequence <400> 78 gacatccaga tgacccagtc tccttcttct ctgtctgcct ctgtgggcga gagagtgacc 60 atcacctgta aggcctctga ggacatctac aacagactgg cctggtacca gcagaagcct 120 ggcaaggccc ctaagctgct gctgtacggc gagacaggcc tggaggccgg cttccacctg 180 agattcagcg gctctggctc tggcaaacac tacaccctga ccatctcttc tctgcagcct 240 gaggatttcg ccacctacta ctgtcagcag tactggtcta ccccttggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 79 <211> 321 <212> DNA <213> Artificial sequence <400> 79 gacatccaga tgacccagtc ttcttcttct ctgtctgcct ctgtgggcga gagagtgacc 60 atcacctgta aggcctctga ggacatctac aacagactgg cctggtacca gcagaagcct 120 ggcaaggccc ctaagctgct gctgtacggc gagacaggcc tggaggccgg cttccacctg 180 agattcagcg gctctggctc tggcacccac tacaccctga ccatctcttc tctgcagcct 240 gaggatttcg ccacctacta ctgtcagcag tactggtcta ccccttggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 80 <211> 321 <212> DNA <213> Artificial sequence <400> 80 gacatccaga tgacccagtc tccttcttct ctgtctgcct ctgtgggcga gagagtgacc 60 atcacctgta aggcctctga ggacatctac aacagactgg cctggtacca gcagaagcct 120 ggcaaggccc ctaagctgct gctgtacggc gagacaggcc tggaggccgg cttccacctg 180 agattcagcg gctctggctc tggcacccac tacaccctga ccatctcttc tctgcagact 240 gaggatttcg ccacctacta ctgtcagcag tactggtcta ccccttggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 81 <211> 321 <212> DNA <213> Artificial sequence <400> 81 gacatccaga tgacccagtc tccttcttct ctgtctgcct ctgtgggcga gagagtgacc 60 atcacctgta aggcctctga ggacatctac aacagactgg cctggtacca gcagaagcct 120 ggcaacgccc ctaagctgct gctgtacggc gagacaggcc tggaggccgg cttccacctg 180 agattcagcg gctctggctc tggcacccac tacaccctga ccatctcttc tctgcagcct 240 gaggatttcg ccacctacta ctgtcagcag tactggtcta ccccttggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 82 <211> 321 <212> DNA <213> Artificial sequence <400> 82 gacatccaga tgacccagtc tccttcttct ctgtctgcct ctgtgggcga gagagtgacc 60 atcacctgta aggcctctga ggacatctac aacagactgg cctggtacca gcagaagcct 120 ggcaaggccc ctaagctgct gctgtacggc gagacaggcc tggaggccgg cttccacctg 180 agattcagcg gctctggctc tggcaccgac tacaccctga ccatctcttc tctgcagcct 240 gaggatttcg ccacctacta ctgtcagcag tactggtcta ccccttggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 83 <211> 321 <212> DNA <213> Artificial sequence <400> 83 gacatccaga tgacccagtc tccttcttct ctgtctgcct ctgtgggcga gagagtgacc 60 atcacctgta aggcctctga ggacatctac aacagactgg cctggtacca gcagaagcct 120 ggcaaggccc ctaagctgct gctgtacggc gagacaggcc tggaggccgg cttccacctg 18

END

END

END

Claims

1. An antibody or antigen-binding fragment that binds to human plasma prokallikrein, characterized in that, The antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region includes the following three complementarity-determining regions (CDRs): CDR1 sequence is DYEMH, CDR2 sequence is AIHPGSGGTAYNQKFKG, and CDR3 sequence is PYGYPFAY; the light chain variable region includes the following three complementarity-determining regions (CDRs): CDR1 sequence is KASEDIYNRLA, CDR2 sequence is GETGLEA, and CDR3 sequence is QQYWSTPWT.

2. The antibody or antigen-binding fragment according to claim 1, characterized in that, The antibody or antigen-binding fragment is a murine antibody, chimeric antibody, humanized antibody, or fully humanized antibody or its antigen-binding fragment.

3. The antibody or antigen-binding fragment according to claim 1, wherein: The heavy chain variable region sequence is SEQ ID NO: 1, and the light chain variable region sequence is selected from any one of SEQ ID NO: 24-32; The heavy chain variable region sequence is SEQ ID NO: 2, and the light chain variable region sequence is selected from any one of SEQ ID NO: 24-26, 28 or 31; The heavy chain variable region sequence is SEQ ID NO: 3, and the light chain variable region sequence is selected from any one of SEQ ID NO: 24-26, 28 or 31; The heavy chain variable region sequence is SEQ ID NO: 4, and the light chain variable region sequence is selected from any one of SEQ ID NO: 24-26, 28; The heavy chain variable region sequence is SEQ ID NO: 5, and the light chain variable region sequence is selected from SEQ ID NO: 24; The heavy chain variable region sequence is SEQ ID NO: 6, and the light chain variable region sequence is selected from any one of SEQ ID NO: 24-26, 28; The heavy chain variable region sequence is SEQ ID NO: 7, and the light chain variable region sequence is selected from SEQ ID NO: 24; The heavy chain variable region sequence is SEQ ID NO: 8, and the light chain variable region sequence is selected from SEQ ID NO: 24; The heavy chain variable region sequence is SEQ ID NO: 9, and the light chain variable region sequence is selected from any one of SEQ ID NO: 33-35; The heavy chain variable region sequence is SEQ ID NO: 10, and the light chain variable region sequence is selected from any one of SEQ ID NO: 33-35; The heavy chain variable region sequence is SEQ ID NO: 11, and the light chain variable region sequence is selected from any one of SEQ ID NO: 33-35; The heavy chain variable region sequence is SEQ ID NO: 12, and the light chain variable region sequence is selected from any one of SEQ ID NO: 33-35 and 42; The heavy chain variable region sequence is SEQ ID NO: 13, and the light chain variable region sequence is selected from any one of SEQ ID NO: 33-35; The heavy chain variable region sequence is SEQ ID NO: 15, and the light chain variable region sequence is selected from any one of SEQ ID NO: 36-42 and 25; The heavy chain variable region sequence is SEQ ID NO: 16, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 17, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 18, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 19, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 20, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 21, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 22, and the light chain variable region sequence is selected from SEQ ID NO: 42; The heavy chain variable region sequence is SEQ ID NO: 23, and the light chain variable region sequence is selected from SEQ ID NO:

42.

4. The antibody or antigen-binding fragment according to claim 3, characterized in that, The antibody heavy chain amino acid sequence is as shown in SEQ ID NO: 89, and the light chain amino acid sequence is as shown in SEQ ID NO: 91; or the antibody heavy chain nucleotide sequence is as shown in SEQ ID NO: 90, and the light chain nucleotide sequence is as shown in SEQ ID NO:

92.

5. The antibody or antigen-binding fragment according to any one of claims 1-4, characterized in that, The antigen-binding fragment is selected from Fab, Fab', Fv, scFv, or (Fab')2 fragments.

6. An isolated nucleic acid that encodes an antibody or antigen-binding fragment as described in any one of claims 1-5.

7. A vector comprising the nucleic acid of claim 6, wherein the vector is an expression vector.

8. A host cell comprising the nucleic acid of claim 6 or the vector of claim 7, wherein the host cell is a prokaryotic cell or a eukaryotic cell.

9. The host cell according to claim 8, characterized in that, The host cells are selected from yeast cells and mammalian cells.

10. A pharmaceutical composition, characterized in that... It comprises an antibody or antigen-binding fragment as described in any one of claims 1-5, and a pharmaceutically acceptable excipient.

11. The pharmaceutical composition according to claim 10, characterized in that, The pharmaceutical composition is an intravitreal injection formulation.

12. Use of the antibody or antigen-binding fragment of any one of claims 1-5 in the preparation of a medicament for treating diabetic macular edema.

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