Compositions comprising anti-cd300c monoclonal antibodies for preventing or treating cancer

Abstract

The present application relates to an anti-CD300c monoclonal antibody, a composition for preventing or treating cancer comprising the antibody and a composition for anti-cancer immunotherapy comprising the antibody, the anti-CD300c monoclonal antibody of the present application not only binds to the CD300c antigen with high specificity, but also can promote anti-cancer immune effects, and is expected to be effectively used for inhibiting the growth, development and metastasis of various cancers.

Description

Technical Field

[0001] This invention relates to an anti-CD300c monoclonal antibody, a composition comprising the antibody for the prevention or treatment of cancer, and a composition comprising the antibody for anti-cancer immunotherapy. Background Technology

[0002] Cancer is one of the leading causes of death in modern society. It is a disease caused by gene mutations resulting from various factors, leading to changes in normal cells. It refers to malignant tumors that deviate from normal cell differentiation, proliferation, and growth patterns. Cancer is characterized by "uncontrolled cell growth," where abnormal cell growth forms clusters of cells called tumors that infiltrate surrounding tissues and, in severe cases, metastasize to other organs. Cancer is a difficult-to-treat chronic disease; even with treatments such as surgery, radiation, and drug therapy, it is often incurable, causing immense suffering and ultimately leading to death. In recent years, due to factors such as an aging population and environmental degradation, the global cancer incidence rate has been increasing by more than 5% annually. According to the World Health Organization (WHO), it is estimated that within the next 25 years, the number of people diagnosed with cancer will increase to 30 million, with 20 million dying from it.

[0003] Cancer drug therapy, or anticancer drugs, is typically a cytotoxic compound that treats cancer by attacking and causing cancer cells to die. However, it damages not only cancer cells but also normal cells, resulting in significant side effects. Therefore, targeted anticancer drugs have been developed to reduce these side effects. However, while these targeted anticancer drugs can reduce side effects, they also have the limitation of a high probability of developing drug resistance. Therefore, there has been a surge in interest in immunotherapy drugs that utilize the body's immune system to reduce problems caused by toxicity and drug resistance. As an example of such immunotherapy drugs, immune checkpoint inhibitors have been developed. These specifically bind to PD-L1 on the surface of cancer cells to inhibit the binding of T cells to PD-1, thereby activating T cells and enabling them to attack cancer cells (Korean Patent Publication No. 10-2018-0099557). However, even with these immune checkpoint inhibitors, the number of cancer types that are effective is limited. Therefore, there is an urgent need to develop novel anticancer immunotherapy drugs that can produce the same therapeutic effect on multiple cancers. Summary of the Invention

[0004] Technical issues

[0005] The present invention is proposed to solve the problems in the prior art, and its purpose is to provide a composition comprising an anti-CD300c monoclonal antibody for the prevention or treatment of cancer, and a composition comprising the same for anti-cancer immunotherapy.

[0006] However, the problems to be solved by the present invention are not limited to those described above. Those skilled in the art can clearly understand other unmentioned problems through the following description.

[0007] Technical solution

[0008] This invention provides an anti-CD300c monoclonal antibody comprising one or more complementarity-determining region (CDR) sequences selected from the group consisting of sequences 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, and 50. The CDR sequences may include amino acid sequences having at least 90%, more preferably at least 95%, and most preferably at least 98% sequence homology to one or more CDR sequences selected from the group consisting of sequences 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, and 50. The "percentage of sequence homology" of an amino acid sequence is determined by comparing it with an optimally aligned sequence and a comparison region, wherein a portion of the amino acid sequence in the comparison region may include additions or deletions (i.e. gaps) compared with a reference sequence of optimal alignment (excluding additions or deletions).

[0009] In a specific embodiment of the present invention, the above-mentioned anti-CD300c monoclonal antibody may have cross-reactivity. Preferably, the cross-reactivity may refer to the cross-reactivity between human CD300c antigen and mammalian CD300c antigen. More preferably, it may refer to the cross-reactivity between human antigen and mouse antigen.

[0010] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising the above-mentioned anti-CD300c monoclonal antibody as an active ingredient.

[0011] In a specific embodiment of the present invention, the aforementioned cancers are preferably colorectal cancer, rectal cancer, colon cancer, thyroid cancer, oral cancer, pharyngeal cancer, laryngeal cancer, cervical cancer, brain cancer, lung cancer, ovarian cancer, bladder cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer, skin cancer, tongue cancer, breast cancer, uterine cancer, stomach cancer, bone cancer, and leukemia, etc., but are not limited to any cancer that expresses CD300c protein on the surface of cancer cells.

[0012] In another specific embodiment of the present invention, the above-mentioned pharmaceutical composition may further comprise other existing immunotherapeutic drugs or anticancer drugs, wherein the immunotherapeutic drugs are preferably anti-PD-1, anti-PD-L1, anti-CTLA-4, anti-KIR, anti-LAG3, anti-CD137, anti-OX40, anti-CD276, anti-CD27, or anti-G ITR (anti-GITR), anti-TIM3 (anti-TIM3), anti-41BB (anti-41BB), anti-CD226 (anti-CD226), anti-CD40 (anti-CD40), anti-CD70 (anti-CD70), anti-ICOS (anti-ICOS), anti-CD40L (anti-CD40L), anti-BTLA (anti-BTLA), anti-TCR (anti-TCR), and anti-TIGIT (anti-TIGIT), etc., but not limited to any substance currently used as an immunotherapy drug. Furthermore, the preferred anticancer drugs are doxorubicin, cisplatin, gemcitabine, oxaliplatin, 5-FU, cetuximab, panitumumab, nimotuzumab, necitumumab, cancer antigens, anticancer viruses, etc., but not limited to any substance currently used as an anticancer drug. The aforementioned cancer antigens are cancer vaccines specific to certain cancer types. For bladder cancer, NY-ESO-1 is preferred; for breast cancer, HER2 is preferred; for colorectal cancer, CEA is preferred; and for lung cancer, VEGFR1 and VEGFR2 are preferred. However, any known cancer antigen used as a cancer vaccine is not limited to these. Examples of anticancer viruses include Imlygic and Pexa-Vec, but any known anticancer virus is not limited to these. The aforementioned anticancer drugs can preferably be used in combination with the monoclonal antibody disclosed herein, or they can be in a form bound to the monoclonal antibody of this invention, or they can be co-encapsulated within an anticancer drug carrier.

[0013] In another specific embodiment of the present invention, the above-mentioned pharmaceutical composition is characterized by inhibiting the proliferation, survival, metastasis, recurrence, and drug resistance of cancer or cancer stem cells, but the effects produced by the pharmaceutical composition of the present invention are not limited thereto.

[0014] Furthermore, the present invention provides an immunotherapeutic drug comprising the above-mentioned anti-CD300c monoclonal antibody as an active ingredient.

[0015] Furthermore, the present invention provides an adjuvant for anticancer therapy, which comprises the above-mentioned anti-CD300c monoclonal antibody as an active ingredient.

[0016] In one specific embodiment of the present invention, the above-mentioned adjuvant can enhance the anti-cancer treatment effect by activating the immune function of immune cells.

[0017] In another specific embodiment of the present invention, the above-mentioned anticancer therapy may be radiotherapy, anticancer drug therapy or immunotherapy.

[0018] Furthermore, the present invention provides a cancer treatment method comprising the step of administering to an individual a composition containing the above-described anti-CD300c monoclonal antibody as an active ingredient.

[0019] Furthermore, the present invention provides the use of a composition comprising the above-mentioned anti-CD300c monoclonal antibody as an active ingredient for the prevention or treatment of cancer.

[0020] Furthermore, the present invention provides the use of the above-mentioned anti-CD300c monoclonal antibody to prepare a drug for cancer treatment.

[0021] The effects of the invention

[0022] The anti-CD300c monoclonal antibody of this invention binds specifically to CD300c expressed on the surface of various cancer cells with high binding affinity, thereby activating T cells and promoting differentiation into M1 macrophages. Therefore, it can effectively inhibit cancer cell proliferation and can be effectively used as an immunotherapy drug for various cancers. Furthermore, the anti-CD300c monoclonal antibody of this invention can not only enhance the therapeutic effect by combining with existing immunotherapeutic drugs, but also exhibits cross-species cross-reactivity, thus allowing for wide application in various mammals. Moreover, treatment of apoptosis-resistant cancer cells with the anti-CD300c monoclonal antibody of this invention can significantly weaken the cancer cell resistance, showing promising efficacy in preventing cancer recurrence. Furthermore, cancer cells often evade the immune system by inhibiting the production of the proinflammatory cytokine interleukin-2 (IL-2), and it has been confirmed that the anti-CD300c monoclonal antibody can restore the production of IL-2 blocked by these cancer cells, inducing apoptosis by activating the immune system, thus allowing it to be used as a more fundamental immunotherapeutic drug. Attached Figure Description

[0023] Figure 1 A simplified diagram illustrating the mechanism by which the anti-cancer effects of the anti-CD300c monoclonal antibody and / or CD300c small interfering RNA (siRNA) of the present invention are generated.

[0024] Figure 2 A simplified diagram illustrating the mechanism of action of the anti-CD300c monoclonal antibody of the present invention on various monocytes, T cells, and cancer cells.

[0025] Figure 3 This is an SDS-PAGE result of an anti-CD300c monoclonal antibody under non-reducing conditions according to an embodiment of the present invention.

[0026] Figure 4 This is an SDS-PAGE result of an anti-CD300c monoclonal antibody under reducing conditions according to an embodiment of the present invention.

[0027] Figure 5 The diagram shows the results confirming the binding affinity of an anti-CD300c monoclonal antibody to the CD300c antigen according to an embodiment of the present invention.

[0028] Figure 6 The figure shows the results of confirming the T-cell activation ability of the anti-CD300c monoclonal antibody of an embodiment of the present invention by enzyme-linked immunosorbent assay (ELISA).

[0029] Figure 7 The figure shows the results of ELISA confirmation of the differentiation ability of the anti-CD300c monoclonal antibody of an embodiment of the present invention into M1 macrophages.

[0030] Figure 8 The figure shows the results of ELISA confirmation of the differentiation ability of the anti-CD300c monoclonal antibody of an embodiment of the present invention into M1 macrophages.

[0031] Figure 9 The figure shows the results of ELISA confirming the differentiation ability of the anti-CD300c monoclonal antibody of an embodiment of the present invention into M1 macrophages at different concentrations.

[0032] Figure 10 The figure shows the results of ELISA confirming the differentiation ability of the anti-CD300c monoclonal antibody of an embodiment of the present invention into M1 macrophages at different concentrations.

[0033] Figure 11 This diagram illustrates the results of confirming the differentiation ability of an anti-CD300c monoclonal antibody of an embodiment of the present invention into M1 macrophages through cell morphology.

[0034] Figure 12 The figure shows the results of ELISA confirmation of the differentiation ability of the CL7 anti-CD300c monoclonal antibody of an embodiment of the present invention into M1 macrophages.

[0035] Figure 13 This figure illustrates the results of comparing the differentiation ability of an anti-CD300c monoclonal antibody and an immunotherapy drug into M1 macrophages using ELISA, according to an embodiment of the present invention.

[0036] Figure 14 The figure shows the results of comparing the differentiation ability of the CL7 anti-CD300c monoclonal antibody and the immunotherapy drug of an embodiment of the present invention into M1 macrophages by ELISA.

[0037] Figure 15 The figure shows the results of comparing the differentiation ability of the CL7 anti-CD300c monoclonal antibody and the immunotherapy drug of an embodiment of the present invention into M1 macrophages by ELISA.

[0038] Figure 16 The figure shows the results of comparing the differentiation ability of the CL7 anti-CD300c monoclonal antibody and the immunotherapy drug of an embodiment of the present invention into M1 macrophages by ELISA.

[0039] Figure 17 The figure shows the results of comparing the differentiation ability of M0 macrophages into M1 macrophages by ELISA of the CL7 anti-CD300c monoclonal antibody and the immunotherapeutic drug according to an embodiment of the present invention.

[0040] Figure 18 This figure illustrates the results of comparing the differentiation ability of an anti-CD300c monoclonal antibody and an immunotherapy drug into M1 macrophages using ELISA, according to an embodiment of the present invention.

[0041] Figure 19 The figure shows the results of ELISA confirming the ability of an anti-CD300c monoclonal antibody according to an embodiment of the present invention to redifferentiate M2 macrophages into M1 macrophages.

[0042] Figure 20 The figure shows the results of ELISA confirming the ability of an anti-CD300c monoclonal antibody according to an embodiment of the present invention to redifferentiate M2 macrophages into M1 macrophages.

[0043] Figure 21 The figure shows the results of ELISA confirming the ability of an anti-CD300c monoclonal antibody according to an embodiment of the present invention to redifferentiate M2 macrophages into M1 macrophages.

[0044] Figure 22 The figure shows the results of ELISA confirmation of the ability of the anti-CD300c monoclonal antibody of an embodiment of the present invention to redifferentiate M0, M1 and M2 macrophages into M1 macrophages.

[0045] Figure 23 The figure shows the results of combining an anti-CD300c monoclonal antibody with an anti-PD-L1 immunotherapy drug to demonstrate the drug's ability to differentiate into M1 macrophages, as described in an embodiment of the present invention.

[0046] Figure 24 The figure shows the results of combining an anti-CD300c monoclonal antibody with an immunotherapeutic drug to demonstrate the ability of the antibody to differentiate into M1 macrophages, as described in an embodiment of the present invention.

[0047] Figure 25 The figure shows the results confirming the inhibitory effect of an anti-CD300c monoclonal antibody of an embodiment of the present invention on cancer cell growth under 0% fetal bovine serum (FBS) conditions.

[0048] Figure 26 The figure shows the results confirming the inhibitory effect of the anti-CD300c monoclonal antibody of an embodiment of the present invention on cancer cell growth under 0.1% FBS conditions.

[0049] Figure 27 The figure shows the results of comparing the inhibitory effects of an anti-CD300c monoclonal antibody and an immunotherapeutic drug on the growth of cancer cells (lung cancer) according to an embodiment of the present invention.

[0050] Figure 28 The figure shows the results of comparing the inhibitory effects of an anti-CD300c monoclonal antibody and an immunotherapeutic drug on the growth of cancer cells (breast cancer) according to an embodiment of the present invention.

[0051] Figure 29 The figure shows the results confirming the inhibitory effect of the anti-CD300c monoclonal antibody of an embodiment of the present invention on cancer cell growth at different concentrations.

[0052] Figure 30 The figure shows the results confirming the inhibitory effect of the combined use of an anti-CD300c monoclonal antibody and an immunotherapy drug on cancer cell growth according to an embodiment of the present invention.

[0053] Figure 31 The figure shows the results confirming the inhibitory effect of the combined use of an anti-CD300c monoclonal antibody and an immunotherapy drug on cancer cell growth according to an embodiment of the present invention.

[0054] Figure 32 The diagram illustrates the results confirming the mechanism of action of the combined use of an anti-CD300c monoclonal antibody and an immunotherapeutic drug according to an embodiment of the present invention.

[0055] Figure 33 The diagram shows the results confirming the binding specificity of the anti-CD300c monoclonal antibody according to an embodiment of the present invention.

[0056] Figure 34 The diagram shows the results confirming the cross-reactivity of an anti-CD300c monoclonal antibody according to an embodiment of the present invention in mice.

[0057] Figure 35 The image shows the results confirming the anti-cancer effect (colorectal cancer) of an anti-CD300c monoclonal antibody according to an embodiment of the present invention on mice.

[0058] Figure 36 The diagram illustrates the results confirming the effect of an anti-CD300c monoclonal antibody of an embodiment of the present invention on cancer growth in vivo.

[0059] Figure 37 The figure shows the results confirming the inhibitory effect of an anti-CD300c monoclonal antibody of an embodiment of the present invention on cancer growth in vivo.

[0060] Figure 38 This is a graph illustrating the effect of an anti-CD300c monoclonal antibody, according to an embodiment of the present invention, on increasing tumor-infiltrating lymphocytes in the in vivo tumor microenvironment. Scale bars represent 50 μm.

[0061] Figure 39 The figure shows the results confirming the effect of an anti-CD300c monoclonal antibody according to an embodiment of the present invention on the increase of M1 macrophages in vivo. Detailed Implementation

[0062] The anti-CD300c monoclonal antibody of the present invention effectively inhibits CD300c by specifically binding to the CD300c protein with high binding affinity, thereby activating T cells. It can also effectively inhibit the growth of cancer cells, cancer development and metastasis by promoting differentiation into M1 macrophages. Therefore, it can be effectively used to treat various cancers that express CD300c antigen on their surface.

[0063] In this specification, the term "antibody" refers to an immunoglobulin molecule that immunologically reacts with a specific antigen, including polyclonal antibodies, monoclonal antibodies, and their functional fragments. Furthermore, the term can include genetically engineered forms such as chimeric antibodies (e.g., humanized mouse antibodies) and heterologous binding antibodies (e.g., bispecific antibodies). Monoclonal antibodies, unlike polyclonal antibodies which contain multiple antibodies specific to different epitopes, exhibit binding specificity and affinity only to a single epitope on the antigen, thus making it easier to control the quality of therapeutic agents. In particular, the anti-CD300c monoclonal antibody of the present invention not only specifically binds to cancer cells expressing CD300c, thereby exhibiting anticancer activity itself, but also maximizes cancer cell-dependent anticancer activity by stimulating immune cells. The structure of the antibody described above includes a variable region of heavy chain and / or light chain. The variable region includes the portion that forms the antigen-binding site of the antibody molecule as a primary structure. The antibody of the present invention can be composed of some fragments containing the above variable region. Preferably, the variable region can be replaced by a soluble receptor of CD300c, but it is not limited thereto as long as it can produce the same effect as the anti-CD300c monoclonal antibody of the present invention.

[0064] In this specification, the term "immunoglobulin" is a concept that includes both antibody-like molecules and antibodies that have the same structural characteristics as antibodies but do not have antigen specificity.

[0065] In this specification, the term "single-chain variable fragment (scFv)" refers to a protein in which the variable regions of the light and heavy chains of an antibody are linked by a linker consisting of a peptide sequence of approximately 15 amino acids. This linker can have a light chain variable region-linker-heavy chain variable region or heavy chain variable region-linker-light chain variable region sequence, and possesses the same or similar antigen specificity as the original antibody. The linker site is a hydrophilic, flexible peptide chain primarily composed of glycine and serine, mainly using the 15-amino acid sequence "(Gly-Gly-Gly-Ser)3" or a similar sequence.

[0066] In this specification, the term "immunotherapy drugs" refers to a general term for cancer treatment methods that can activate the immune function of immune cells in the body to fight cancer cells. Examples include, but are not limited to, anti-PD-1, anti-PD-L1, anti-CTLA-4, anti-KIR, anti-LAG3, anti-CD137, anti-OX40, anti-CD276, anti-CD27, anti-GITR, anti-TIM3, anti-41BB, anti-CD226, anti-CD40, anti-CD70, anti-ICOS, anti-CD40L, anti-BTLA, anti-TCR, and anti-TIGIT.

[0067] In this specification, the term "adjuvant" refers to an adjuvant used to improve and / or enhance the therapeutic effect by assisting the efficacy of the main drug, i.e., the anticancer drug, or to improve and / or enhance the therapeutic effect by inhibiting drug resistance to the main drug, or for the purpose of preventing or mitigating the adverse effects of the main drug. The adjuvants of the present invention are not limited as long as they contain anti-CD300c monoclonal antibodies.

[0068] In this specification, the term "prevention" refers to any action that inhibits or delays the onset of diseases such as cancer by applying the pharmaceutical composition of the present invention.

[0069] In this specification, the term "treatment" means any action that improves or brings about a beneficial change in the symptoms of cancer or the like by applying the pharmaceutical composition of the present invention.

[0070] In this specification, the term "individual" refers to any object to which the pharmaceutical composition of the present invention may be administered, and there are no limitations on the subject.

[0071] In this specification, the term "pharmaceutical composition" can be characterized as capsules, tablets, granules, injections, ointments, powders, or beverages, and the pharmaceutical composition can be designed for human use. The pharmaceutical composition is not limited to these forms and can be prepared into oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, as well as external dosage forms, suppositories, and sterile injectable solutions using conventional methods. The pharmaceutical composition of the present invention can contain a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include, for oral administration, binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, and fragrances; for injections, buffers, preservatives, analgesics, solubilizers, isotonic agents, and stabilizers can be mixed in; and for topical administration, bases, excipients, lubricants, and preservatives can be used. The dosage forms of the pharmaceutical composition of the present invention can be prepared in various forms by mixing with the aforementioned pharmaceutically acceptable carriers. For example, when administered orally, it can be formulated as tablets, lozenges, capsules, elixirs, suspensions, syrups, crystal tablets, etc.; when used as an injectable, it can be formulated as single-dose ampoules or multiple-dose formulations. Furthermore, it can be formulated as solutions, suspensions, tablets, capsules, sustained-release formulations, etc.

[0072] Examples of suitable carriers, excipients, and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, or mineral oil. Additionally, fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, and preservatives may also be included.

[0073] The routes of administration of the pharmaceutical compositions according to the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrasheath, intracardiac, percutaneous, subcutaneous, intraperitoneal, intranasal, intestinal, local, sublingual, or rectal administration. Oral or non-oral administration is preferred. The term "parenteral" as used in this application includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrabursal, intrasternal, intrasheath, intralesional, and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention can also be used for rectal administration in suppository form.

[0074] The pharmaceutical compositions of the present invention can vary depending on various factors such as the activity of the specific compound used, age, weight, general health condition, gender, formulation, administration time, route of administration, excretion rate, drug combination, and severity of the specific disease to be prevented or treated. The dosage of the above-mentioned pharmaceutical compositions can be appropriately selected by those skilled in the art based on the patient's condition, weight, disease severity, drug form, route of administration, and cycle. The daily dosage can be from 0.0001 mg / kg to 500 mg / kg or from 0.001 mg / kg to 500 mg / kg. It can be administered once daily or in divided doses. The above dosages do not limit the scope of the invention in any way. The pharmaceutical compositions of the present invention can be formulated into pills, sugar-coated pills, capsules, liquids, gels, syrups, pastes, and suspensions.

[0075] The following embodiments are provided to facilitate understanding of the present invention. However, the following embodiments are provided only for easier understanding of the present invention, and the content of the present invention is not limited to the following embodiments.

[0076] Example

[0077] Example 1: Screening for anti-CD300c monoclonal antibodies

[0078] 1.1. Preparation of anti-CD300c monoclonal antibody library

[0079] To screen for anti-CD300c monoclonal antibodies, biopanning was performed using λ phage libraries, κ phage libraries, VH3VL1 phage libraries, and OPALTL phage libraries. More specifically, CD300c antigen at a concentration of 5 μg / mL was added to immunotubes and reacted for 1 hour to allow the antigen to adsorb onto the tube surface. Then, after inhibiting nonspecific reactions by adding 3% skim milk, 10 μg / mL of the antigen dispersed in the 3% skim milk was... 12PFU antibody phage libraries were added to each immunoassay tube and bound to the antigen. The tubes were then washed three times with TBST (tris-buffered salt-Tween 20) to remove non-specifically bound phages. Single-chain variable fragment phage antibodies specifically bound to the CD300c antigen were eluted with 100 mM triethylamine. The eluted phages were neutralized with 1.0 M Tris-HCl buffer (pH 7.8) and then *E. coli* ER2537 was infected at 37°C for 1 hour. The infected *E. coli* were plated on LB agar containing carbenicillin and incubated at 37°C for 16 hours. The resulting *E. coli* colonies were then resuspended in 3 mL of superbroth (SB)-carbenicillin broth. A portion of the colonies was added to 15% glycerol and stored at -80°C until use. The remainder was re-inoculated in SB-carbenicillin-2% glucose solution and incubated at 37°C. The obtained culture medium was then centrifuged, and the supernatant containing phage particles was used to perform three more biological panning processes to obtain antigen-specific antibodies, which were then concentrated.

[0080] After three rounds of biological panning, *E. coli* containing the antibody gene were plated on LB agar medium containing carbenicillin and incubated at 37°C for 16 hours. The resulting *E. coli* colonies were then inoculated again into SB-carbenicillin-2% glucose solution and incubated at 37°C until the absorbance (OD) reached a certain level. 600nm After the concentration was reduced to 0.5, isopropyl-β-D-thiogalactoside (IPTG) was added and the mixture was further cultured at 30°C for 16 hours. Subsequently, periplasmic extraction was performed, and the antibody library pool that specifically binds to the CD300c antigen was obtained for the first time based on the above results.

[0081] 1.2. Screening for anti-CD300c monoclonal antibodies

[0082] To screen for anti-CD300c monoclonal antibodies that specifically bind to the CD300c antigen with high binding affinity, ELISA was performed using a library pool obtained using the same method as in Example 1.1. More specifically, CD300c and CD300a antigens were aliquoted into ELISA plates at a concentration of 5 μg / mL per well in coating buffer (0.1 M sodium carbonate, pH 9.0) and incubated at room temperature for 3 hours to bind the antigens to the plates. Then, the plates were washed three times with phosphate-buffered saline-Tween 20 (PBST) to remove unbound antigens. Next, 350 μL of PBST supplemented with 2% bovine serum albumin (BSA) was added to each well, and the plates were incubated at room temperature for 1 hour, followed by washing with PBST again. Then, 25 μg of periplasmic extract containing scFv, obtained using the same method as in Example 1.1, was added to each well, and the plates were incubated at room temperature for 1 hour to bind the antigens. One hour later, the sample was washed three times with PBST to remove unbound scFv. Then, 4 μg / mL of the detection antibody was added, and the reaction was carried out again at room temperature for one hour. Next, after removing the unbound detection antibody with PBST, anti-rabbit IgG conjugated with horseradish peroxidase (HRP) was added, and the reaction was carried out at room temperature for one hour. Unbound antibodies were removed again with PBST. Then, TMB solution was added, and after reacting for 10 minutes for color development, the reaction was terminated by adding 2N sulfuric acid solution. The absorbance was measured at 450 nm to confirm the antibody specifically binding to the CD300c antigen.

[0083] 1.3. Confirmation of the anti-CD300c monoclonal antibody sequence

[0084] The nucleotide sequences of the screened anti-CD300c monoclonal antibodies were confirmed using the same method as in Example 1.2. More specifically, plasmid DNA was extracted from the screened antibody clones using a plasmid miniprepkit, followed by DNA sequencing to analyze the complementarity-determining region (CDR) sequences. As a result, 25 anti-CD300c monoclonal antibodies with different amino acid sequences were obtained.

[0085] Three anti-CD300c monoclonal antibodies were screened using the λ phage library: SL18 (DNA sequence 35) containing the CDR sequence of sequence 36, CL4 (DNA sequence 7) containing the CDR sequence of sequence 8, and CL5 (DNA sequence 9) containing the CDR sequence of sequence 10.

[0086] Ten anti-CD300c monoclonal antibodies were screened using the κ phage library: CK1 (DNA sequence: sequence 1) containing the CDR sequence of sequence 2, CK2 (DNA sequence: sequence 3) containing the CDR sequence of sequence 4, CK3 (DNA sequence: sequence 5) containing the CDR sequence of sequence 6, SK11 (DNA sequence: sequence 21) containing the CDR sequence of sequence 22, SK12 (DNA sequence: sequence 23) containing the CDR sequence of sequence 24, SK13 (DNA sequence: sequence 25) containing the CDR sequence of sequence 26, SK14 (DNA sequence: sequence 27) containing the CDR sequence of sequence 28, SK15 (DNA sequence: sequence 29) containing the CDR sequence of sequence 30, SK16 (DNA sequence: sequence 31) containing the CDR sequence of sequence 32, and SK17 (DNA sequence: sequence 33) containing the CDR sequence of sequence 34.

[0087] Ten anti-CD300c monoclonal antibodies were screened from the VH3VL1 phage library: CB301_H3L1_A10 (DNA sequence: sequence 38) containing the CDR sequence of sequence 37; CB301_H3L1_A12 (DNA sequence: sequence 39) containing the CDR sequence of sequence 40; CL6 (DNA sequence: sequence 11) containing the CDR sequence of sequence 12; CB301_H3L1_E6 (DNA sequence: sequence 41) containing the CDR sequence of sequence 42; and [the remaining text appears to be incomplete and requires further context for accurate translation.] CL7 (DNA sequence is sequence 13) contains the CDR sequence 44, CB301_H3L1_F4 (DNA sequence is sequence 43) contains the CDR sequence 16, CL8 (DNA sequence is sequence 15) contains the CDR sequence 16, CB301_H3L1_G11 (DNA sequence is sequence 45) contains the CDR sequence 46, CL9 (DNA sequence is sequence 17) contains the CDR sequence 18, and CL10 (DNA sequence is sequence 19) contains the CDR sequence 20.

[0088] Two anti-CD300c monoclonal antibodies were screened using the OPALTL phage library: CB301_OPALTL_B5 (DNA sequence is sequence 47), which contains the CDR sequence of sequence 48, and CB301_OPALTL_E6 (DNA sequence is sequence 49), which contains the CDR sequence of sequence 50.

[0089] Based on the above results, 25 anti-CD300c monoclonal antibodies with high binding affinity to CD300c antigen were identified that can be used for the prevention or treatment of cancer.

[0090] 1.4. Preparation and purification of anti-CD300c monoclonal antibody

[0091] Using the base sequence of the anti-CD300c monoclonal antibody confirmed in Example 1.3, an expression vector capable of expressing the antibody and with separate heavy and light chains was prepared. More specifically, the vector was prepared by inserting a gene using the analyzed CDR sequence to express the heavy and light chains separately in the pCIW3.3 vector. The prepared heavy and light chain expression vectors were mixed with polyethyleneimine (PEI) at a 1:1 mass ratio and transfected into 293T cells to induce antibody expression. On day 8, the culture medium was centrifuged to remove the cells, yielding the culture medium. The obtained culture medium was filtered and resuspended in a solution of 0.1M NaH2PO4 and 0.1M Na2HPO4 (pH 7.0). The resuspended solution was then purified by affinity chromatography using protein A beads (GE Healthcare), and finally eluted with elution buffer (Thermofisher).

[0092] To confirm the prepared antibody, 5 μg of purified antibody was added to reducing sample buffer and non-reducing sample buffer, respectively, and then electrophoresis was performed using pre-made sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE, Invitrogen). The proteins were then stained with Coomassie Brilliant Blue. Results under each non-reducing condition are shown in the table below. Figure 3 The results under the reduction conditions are shown in the figure. Figure 4 As shown in the image.

[0093] like Figure 3 and Figure 4 As shown, anti-CD300c monoclonal antibody with a purity of over 90% has been prepared and purified.

[0094] 1.5. Confirm the antigen-binding affinity of the anti-CD300c monoclonal antibody.

[0095] To screen for monoclonal antibodies that specifically bind to the CD300c antigen with superior binding affinity from anti-CD300c monoclonal antibodies prepared using the same method as in Example 1.4, a binding affinity enzyme-linked immunosorbent assay (BINDI) was performed. More specifically, CD300c or CD300a antigen was aliquoted into ELISA plates at a concentration of 8 μg / mL per well in coating buffer (0.1 M sodium carbonate, pH 9.0) and incubated at room temperature for 3 hours to bind the antigen. The plates were then washed three times with PBST to remove unbound antigen. 300 μL of PBST supplemented with 5% bovine serum albumin was added to each well, and the plates were incubated at room temperature for 1 hour, followed by another wash with PBST. Then, a quadruplicate of the anti-CD300c monoclonal antibody was added, and the plates were incubated at room temperature for 1 hour to bind the antigen. One hour later, the sample was washed three times with PBST to remove unbound anti-CD300c monoclonal antibody. Then, 4 μg / mL of the detection antibody (HRP-conjugated anti-Fc IgG) was added, and the reaction was repeated at room temperature for one hour. Next, unbound detection antibody was removed with PBST, and TMB solution was added. After reacting for 10 minutes for color development, 2N sulfuric acid solution was added to terminate the reaction. The absorbance was measured at 450 nm to confirm the presence of antibody specifically binding to the CD300c antigen. The results are shown in Table 1 and... Figure 5 As shown in the image.

[0096] Table 1

[0097]

[0098]

[0099] As shown in Table 1, the EC50 (half-maximum effective concentration of drug that causes 50% of the maximum response) of the anti-CD300c monoclonal antibody was determined. Except for four clones (CK3, CL8, SK15, and SK16), the remaining 14 clones were all below 0.2 μg / mL, confirming high binding affinity.

[0100] like Figure 5As shown, the sigmoid curve in the Binding ELISA results also confirms that the anti-CD300c monoclonal antibody of the present invention binds to the CD300c antigen with strong binding force.

[0101] Example 2: Confirmation of the anti-cancer effect of anti-CD300c monoclonal antibody in T cells

[0102] To confirm whether the anti-CD300c monoclonal antibody screened using the method in Example 1.5 could produce an anti-cancer effect by activating T cells, the production level of interleukin-2 (IL-2) was determined. IL-2 is an immune factor that promotes T cell growth, proliferation, and differentiation. Increased IL-2 production indicates increased stimulation that induces T cell differentiation, proliferation, and growth, thereby activating T cells. More specifically, anti-CD3 monoclonal antibody and anti-CD28 monoclonal antibody were added to 96-well plates at a concentration of 2 μg / well and fixed for 24 hours, while simultaneously using 1×10⁻⁶... 5 Human T lymphocytes (Jurkat T) line per cell / well were treated with 10 μg / well of anti-CD300c monoclonal antibody. IL-2 production was then measured using an ELISA kit (IL-2 Quantikine kit, R&D systems) and compared with a control group not treated with anti-CD300c monoclonal antibody. Results were... Figure 6 As shown in the image.

[0103] like Figure 6 As shown, it was confirmed that when Jurkat T cells activated by anti-CD3 and anti-CD28 monoclonal antibodies were treated with anti-CD300c monoclonal antibody, the production of IL-2 increased. The above results confirm that anti-CD300c monoclonal antibody can activate T cells and thereby induce anti-cancer immune effects to inhibit the growth of cancer tissue.

[0104] Example 3: Confirmation of the anticancer effect of anti-CD300c monoclonal antibody on its ability to differentiate into macrophages.

[0105] 3.1. Confirm the differentiation ability of anti-CD300c monoclonal antibody into M1 macrophages.

[0106] To confirm that the anti-CD300c monoclonal antibody screened using the method in Example 1.5 induces monocyte differentiation into M1 macrophages, 1.5 × 10⁻⁶ cells were used. 4Human mononuclear cell line (THP-1) cells / well were aliquoted into 96-well plates and treated with 10 μg / mL anti-CD300c monoclonal antibody and / or 100 ng / mL lipopolysaccharide (LPS). After 48 hours of reaction, the production of tumor necrosis factor-α (TNF-α), a differentiation marker of M1 macrophages, was measured using an ELISA kit (Human TNF-α Quantikine kit, R&D systems). The results were... Figure 7 and Figure 8 As shown in the image.

[0107] like Figure 7 As shown, the production of TNF-α by the anti-CD300c monoclonal antibodies CL4, CL7, CL10, and SL18 was more than twice that of the control group (Con) treated with LPS alone.

[0108] And, as Figure 8 As shown, it was confirmed that the production of TNF-α in the experimental group treated with anti-CD300c monoclonal antibody alone, without LPS treatment, was increased compared with the control group (Con) treated with LPS alone.

[0109] 3.2. Confirm the differentiation capacity into M1 macrophages based on different concentrations of anti-CD300c monoclonal antibody.

[0110] To confirm that the induction of M1 macrophage differentiation by anti-CD300c monoclonal antibody increases with the concentration of anti-CD300c monoclonal antibody, the production of TNF-α was determined using the same method as in Example 3.1. Treatments were performed with anti-CD300c monoclonal antibody at concentrations of 10 μg / mL, 1 μg / mL, and 0.1 μg / mL. The results were... Figure 9 As shown in the image.

[0111] like Figure 9 As shown, it was confirmed that the amount of TNF-α produced increased with the increase of the concentration of anti-CD300c monoclonal antibody.

[0112] To confirm more specific concentrations, treatment with CL7 anti-CD300c monoclonal antibody at concentrations of 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.625 μg / mL, 0.313 μg / mL, 0.157 μg / mL, and 0.079 μg / mL was performed to determine the amount of TNF-α produced. The results were... Figure 10 As shown in the image.

[0113] like Figure 10As shown, it was confirmed that the increase in TNF-α production was dependent on the concentration of anti-CD300c monoclonal antibody.

[0114] 3.3. Confirmation of M1 macrophage differentiation induced by anti-CD300c monoclonal antibody through cell morphology.

[0115] To confirm the differentiation of monocytes into M1 macrophages upon treatment with anti-CD300c monoclonal antibody via cell morphology, THP-1 cells were treated with 10 μg / mL of anti-CD300c monoclonal antibody and cultured for 48 hours before cell morphology was observed under a microscope. The results were... Figure 11 As shown in the image.

[0116] like Figure 11 As shown, the morphology of THP-1 cells in the experimental group (CL7) treated with anti-CD300c monoclonal antibody was confirmed to change from suspension cells to M1 macrophages, i.e., round, attached cells. These results confirm that treatment with anti-CD300c monoclonal antibody can promote the differentiation of monocytes into M1 macrophages.

[0117] 3.4. Confirm the differentiation ability of CL7 anti-CD300c monoclonal antibody into M1 macrophages.

[0118] To further confirm whether the CL7 anti-CD300c monoclonal antibody promotes the differentiation of human monocytes into M1 macrophages, the secretion levels of TNF-α, interleukin-1β (IL-1β), and interleukin-8 (IL-8) were measured using an ELISA kit (R&D systems). More specifically, 1.5 × 10⁻⁶ mmol / L was used. 4 THP-1 cells / well were aliquoted into 96-well plates and treated with 10 μg / mL anti-CD300c monoclonal antibody. After 48 hours of reaction, the production of M1 macrophage differentiation markers TNF-α, IL-1β, and IL-8 was measured using an ELISA kit (Human TNF-α Quantikine kit, R&D systems). The results were... Figure 12 As shown in the image.

[0119] like Figure 12 As shown, it was confirmed that the three types of M1 macrophage differentiation markers were increased in the experimental group treated with anti-CD300c monoclonal antibody compared with the control group (Con) that was not treated with anti-CD300c monoclonal antibody.

[0120] 3.5. Comparison of the differentiation capacity of anti-CD300c monoclonal antibodies and immunotherapeutic drugs into M1 macrophages.

[0121] To compare the differentiation ability of anti-CD300c monoclonal antibodies and immunotherapeutic drugs into M1 macrophages, the production of TNF-α was confirmed using an ELISA kit, following the same method as in Example 3.1. Imfinzi, an anti-PD-L1 immunotherapeutic drug, was used at a concentration of 10 μg / mL. The results were... Figure 13 As shown in the image.

[0122] like Figure 13 As shown, the production of TNF-α by the anti-CD300c monoclonal antibody was significantly increased compared to the control group treated with the known immunotherapeutic drug Imfinzi (Imf) alone. These results confirm that, compared to existing immunotherapeutic drugs, the anti-CD300c monoclonal antibody can significantly enhance the differentiation capacity into M1 macrophages.

[0123] To compare with other immunotherapeutic drugs, patients were treated with 10 μg / mL of the anti-PD-L1 immunotherapeutic drug Imfinzi, the anti-PD-1 immunotherapeutic drug Keytruda, and an Isotype control (immunoglobulin G) antibody, and the production of TNF-α, IL-1β, and IL-8 was confirmed using an ELISA kit. The results were presented in... Figures 14 to 16 As shown in the image.

[0124] like Figures 14 to 16 As shown, it was confirmed that the CL7 anti-CD300c monoclonal antibody significantly increased the production of TNF-α, IL-1β and IL-8 compared with Imfinzi, Keytruda and IgG antibodies. The above results confirm that, compared with existing immunotherapeutic drugs, the anti-CD300c monoclonal antibody can significantly enhance the promotion of differentiation into M1 macrophages.

[0125] 3.6. Comparison of the differentiation capacity of M0 macrophages into M1 macrophages between anti-CD300c monoclonal antibodies and anti-PD-L1 immunotherapeutic drugs.

[0126] To compare the differentiation ability of anti-CD300c monoclonal antibodies and immunotherapeutic drugs into M0 macrophages, 1.5 × 10⁻⁶ cells were used. 4 THP-1 cells / well were aliquoted into 96-well plates and treated with 10 μg / mL anti-CD300c monoclonal antibody, 10 μg / mL Imfinzi, and / or 200 nM propylene glycol methyl ether acetate (PMA, phorbol-12-myristate-13-acetate). After a 48-hour reaction, TNF-α production was measured using an ELISA kit. Results were... Figure 17 As shown in the image.

[0127] like Figure 17 As shown, the control group treated with the immunotherapy drug Imfinzi alone did not produce TNF-α, while the experimental group treated with the anti-CD300c monoclonal antibody alone showed increased TNF-α production. Furthermore, when THP-1 cells were treated with PMA and differentiated into M0 macrophages, the TNF-α production in the anti-CD300c monoclonal antibody-treated group was significantly higher than that in the Imfinzi-treated group. These results confirm that, compared to existing immunotherapy drugs, the anti-CD300c monoclonal antibody is more effective in promoting the differentiation of M0 macrophages into M1 macrophages.

[0128] 3.7. Compare the differentiation capacity of anti-CD300c monoclonal antibodies and anti-PD-L1 immunotherapeutic drugs into M1 macrophages.

[0129] To compare the differentiation ability of anti-CD300c monoclonal antibodies and immunotherapeutic drugs into M1 macrophages, the same method as in Example 3.1 was used to confirm the production of TNF-α. The results were... Figure 18 As shown in the image.

[0130] like Figure 18 As shown, when monocytes were differentiated into M1 macrophages by LPS treatment, there was no significant difference in the production of TNF-α between the experimental groups treated with Imfinzi and LPS simultaneously. However, the production of TNF-α in the experimental group treated with anti-CD300c monoclonal antibody and LPS simultaneously was significantly increased compared with the experimental group treated with anti-CD300c monoclonal antibody alone.

[0131] 3.8. Confirm the ability of anti-CD300c monoclonal antibody to redifferentiate M2 macrophages into M1 macrophages.

[0132] To confirm whether the anti-CD300c monoclonal antibody could redifferentiate M2 macrophages into M1 macrophages, 1.5 × 10⁻⁶ cells were used. 4 THP-1 cells / well were aliquoted into 96-well plates and pretreated with 320 nM PMA for 6 hours. The plates were then treated with 20 ng / mL IL-4 (Interleukin-4) and IL-13 (Interleukin-13) and 10 μg / mL anti-CD300c monoclonal antibody for 18 hours. The production of TNF-α, IL-1β, and IL-8 was then confirmed using an ELISA kit. The results were... Figures 19 to 21 As shown in the image.

[0133] like Figures 19 to 21As shown, it was confirmed that in the experimental group without PMA pretreatment, the production of TNF-α, IL-1β, and IL-8 increased in the experimental group simultaneously treated with IL-4, IL-13, and anti-CD300c monoclonal antibody. Similarly, in the experimental group pretreated with PMA, the production of TNF-α, IL-1β, and IL-8 also increased in the experimental group simultaneously treated with IL-4, IL-13, and anti-CD300c monoclonal antibody. These results confirm that anti-CD300c monoclonal antibody can effectively redifferentiate M2 macrophages into M1 macrophages.

[0134] 3.9. Confirm the ability of anti-CD300c monoclonal antibody to induce the redifferentiation of M0, M1, and M2 macrophages into M1 macrophages.

[0135] To confirm whether the anti-CD300c monoclonal antibody could redifferentiate M0, M1, and M2 macrophages into M1 macrophages, 1.5 × 10⁻⁶ cells were used. 4 THP-1 cells / well were aliquoted into 96-well plates and pretreated with 10 μg / mL anti-CD300c monoclonal antibody for 48 hours. The plates were then treated with 100 ng / mL PMA, 100 ng / mL LPS, and 20 ng / mL IL-4 and IL-13 for 24 hours. TNF-α production was then confirmed using an ELISA kit. The results were... Figure 22 As shown in the image.

[0136] like Figure 22 As shown, the pretreatment groups with anti-CD300c monoclonal antibody confirmed that the production of TNF-α was significantly increased compared with the control groups of M0 macrophages treated with PMA alone, the control groups of M1 macrophages treated with LPS alone, and the control groups of M2 macrophages treated with IL-4 and IL-13 alone. These results confirm that the anti-CD300c monoclonal antibody has excellent ability to differentiate M0, M1, and M2 macrophages into M1 macrophages.

[0137] The above results confirm that, compared with existing immunotherapeutic drugs, anti-CD300c monoclonal antibodies are more effective at promoting differentiation into M1 macrophages, inducing anti-cancer immune responses, and thus inhibiting cancer tissue growth. In particular, it was confirmed that the anti-cancer effect is achieved by redifferentiating M2 macrophages, known to promote cancer cell proliferation and metastasis, into M1 macrophages.

[0138] Example 4: Confirmation of the combined therapeutic effect of anti-CD300c monoclonal antibody and immunotherapeutic drugs

[0139] 4.1. Confirm the efficacy of combined use of anti-CD300c monoclonal antibody and anti-PD-L1 immunotherapy.

[0140] To confirm the therapeutic effect of combining anti-CD300c monoclonal antibody with anti-PD-L1 immunotherapy, the signal transduction of nuclear factor kappa-light-chain enhancer (NF-κB) of activated B cells was identified. More specifically, 8.8 × 10 5 THP-1 cells / well were aliquoted into 6-well plates and treated with 10 μg / mL CL7 anti-CD300c monoclonal antibody and / or 10 μg / mL Imfinzi. After 24 hours of culture, phosphorylated NF-κB (p-NF-κB) was confirmed by Western blotting. The results were... Figure 23 As shown in the image.

[0141] like Figure 23 As shown, the amount of p-NF-κB in the experimental group treated with anti-CD300c monoclonal antibody was increased compared with that treated with the immunotherapeutic drug Imfinzi alone, and the amount of p-NF-κB in the experimental group treated with a combination of anti-CD300c monoclonal antibody and Imfinzi was further increased. These results confirm that the combination of anti-CD300c monoclonal antibody and Imfinzi can promote differentiation into M1 macrophages.

[0142] 4.2. Confirm the efficacy of combined treatment with anti-CD300c monoclonal antibodies, anti-PD-L1 immunotherapy drugs, and / or anti-PD-1 immunotherapy drugs.

[0143] To confirm the efficacy of combined use of anti-CD300c monoclonal antibody, anti-PD-L1 immunotherapy, and / or anti-PD-1 immunotherapy, the signal transduction of p38 mitogen-activated protein kinase (p38 MAPK) and extracellular signal-regulated kinase (ERK) was examined. More specifically, 8.8 × 10⁸ μg / mL of the drug was used. 5 THP-1 cells / well were aliquoted into 6-well plates and treated with 10 μg / mL CL7 anti-CD300c monoclonal antibody, 10 μg / mL Imfinzi, and / or 10 μg / mL Keytruda. After 48 hours of culture, phosphorylated p38 MAPK (p-p38MAPK) and phosphorylated ERK (p-ERK) were confirmed by Western blotting. Results were presented in […]. Figure 24 As shown in the image.

[0144] like Figure 24 As shown, p-p38MAPK and p-ERK proteins were not observed in the experimental groups treated with immunotherapy drugs alone, but both proteins were observed in the group treated with anti-CD300c monoclonal antibody. Furthermore, the amount of p-p38MAPK protein was further increased in the experimental groups treated with combination immunotherapy drugs.

[0145] The above results confirm that anti-CD300c monoclonal antibodies promote differentiation into M1 macrophages through MAPK signaling, and their effects are further enhanced when used in combination with immunotherapy drugs. This confirms that anti-CD300c monoclonal antibodies can be used not only alone as immunotherapy drugs, but also in combination with existing immunotherapy drugs to further enhance the anticancer treatment effect.

[0146] Example 5: Anticancer effect of anti-CD300c monoclonal antibody in vitro

[0147] 5.1. Confirm the inhibitory effect of anti-CD300c monoclonal antibody on cancer cell growth.

[0148] To confirm the effect of the monoclonal antibody targeting CD300c on cancer cell growth, cell proliferation analysis was performed using A549 (human lung cancer cell line). More specifically, 2 × 10⁶ cells were cultured in 0% fetal bovine serum (FBS) and cultured. 4 Cells were aliquoted into 96-well plates, with 6 × 10⁶ cells aliquoted under 0.1% fetal bovine serum conditions. 3 Cells were then treated with 10 μg / mL anti-CD300c monoclonal antibody and cultured for 5 days. Then, they were treated with CCK-8 (DOJINDO) and the OD was measured. 450nm The absorbance at a certain point was used to confirm the inhibitory effect of the anti-CD300c monoclonal antibody on cancer cell growth. The results were... Figure 25 and Figure 26 As shown in the image.

[0149] like Figure 25 As shown, it was confirmed that under 0% FBS conditions, all of them except for the SK11 and SK14 anti-CD300c monoclonal antibodies had the effect of inhibiting cancer cell proliferation.

[0150] like Figure 26 As shown, it was confirmed that all anti-CD300c monoclonal antibodies used in the experiment had the effect of inhibiting cancer cell proliferation under 0.1% FBS conditions.

[0151] 5.2. Comparison of the inhibitory effects of anti-CD300c monoclonal antibodies and immunotherapeutic drugs on cancer cell growth.

[0152] To compare the inhibitory effects of anti-CD300c monoclonal antibodies and immunotherapeutic drugs on cancer cell growth, the inhibitory effects on cell growth were confirmed using A549 (human lung cancer cell line) and MDA-MB-231 (human breast cancer cell line). More specifically, 2 × 10⁻⁶ cells were used under 0% fetal bovine serum conditions. 4 Cells were aliquoted into 96-well plates, with 6 × 10⁶ cells aliquoted under 0.1% fetal bovine serum conditions. 3 Cells were then treated with 10 μg / mL anti-CD300c monoclonal antibody and cultured for 5 days before being observed under a light microscope. The results were... Figure 27 and Figure 28 As shown in the image.

[0153] like Figure 27 As shown, it was confirmed that the anti-CD300c monoclonal antibody can inhibit the proliferation of cancer cells in the A549 cell line more effectively than the immunotherapy drug Imfinzi.

[0154] like Figure 28 As shown, it was confirmed that the anti-CD300c monoclonal antibody can inhibit the proliferation of cancer cells in the MDA-MB-231 cell line more effectively than the immunotherapy drug Imfinzi.

[0155] 5.3. Confirm the inhibitory effect of different concentrations of anti-CD300c monoclonal antibody on cancer cell growth.

[0156] To confirm the inhibitory effect of different concentrations of anti-CD300c monoclonal antibody on cancer cell growth, 2×10⁻⁶ cells were injected into the tumor cells under 0% fetal bovine serum conditions. 4 A549 cells were aliquoted into 96-well plates, treated with 10 μg / mL anti-CD300c monoclonal antibody, and cultured for 5 days. They were then treated with CCK-8 (DOJINDO) and reacted for 3 hours. OD was measured. 450nm The absorbance at a certain point was used to confirm the inhibitory effect of the anti-CD300c monoclonal antibody on cancer cell growth. The results were... Figure 29 As shown in the image.

[0157] like Figure 29 As shown, the inhibitory effect on cancer cell growth was confirmed to be dependent on the concentration of the anti-CD300c monoclonal antibody.

[0158] 5.4. Confirm the therapeutic effect of combined anti-CD300c monoclonal antibody and immunotherapy drugs in cancer treatment.

[0159] To confirm the therapeutic effect of the combination of anti-CD300c monoclonal antibody and immunotherapy drug in cancer treatment, cell proliferation analysis was performed using the same method as in Example 5.1. Imfinzi was used as the immunotherapy drug. The results are in Figure 30 As shown in the image.

[0160] like Figure 30 As shown, it was confirmed that the combination of anti-CD300c monoclonal antibody and immunotherapy drug was more effective in inhibiting cancer cell growth than the immunotherapy drug alone.

[0161] Furthermore, the inhibitory effect on cancer cell growth was confirmed using optical microscopy. The results were... Figure 31 As shown in the image.

[0162] like Figure 31 As shown, it was confirmed that the combination of anti-CD300c monoclonal antibody and immunotherapy drugs effectively inhibited the growth of cancer cells.

[0163] 5.5. Confirm the mechanism of action of combined anti-CD300c monoclonal antibody and immunotherapeutic drugs.

[0164] To confirm the mechanism of action of anti-CD300c monoclonal antibody in combination with immunotherapy in the apoptosis signaling mechanism of cancer cells, A549 cells were treated with 10 μg / mL anti-CD300c monoclonal antibody, Imfinzi, and / or Keytruda, and the amount of cleaved caspase-9, a marker of apoptosis, was confirmed by Western blotting. The results were presented in... Figure 32 As shown in the image.

[0165] like Figure 32 As shown, it has been confirmed that the amount of cleaved Caspase-9 is increased when anti-CD300c monoclonal antibody is used in combination with Imfinzi compared to the experimental group treated with anti-CD300c monoclonal antibody alone. Furthermore, the amount of cleaved Caspase-9 is further increased when anti-CD300c monoclonal antibody, Imfinzi, and Keytruda are used in combination. These results confirm that when anti-CD300c monoclonal antibody is used in combination with immunotherapeutic drugs, the apoptosis signaling mechanism of cancer cells is further activated, thereby effectively inhibiting cancer cell proliferation.

[0166] Example 6: Confirmation of the excellent cross-reactivity of the anti-CD300c monoclonal antibody between human and mouse antigens.

[0167] 6.1. Confirm the specificity of the anti-CD300c monoclonal antibody.

[0168] To confirm the specificity of the anti-CD300c monoclonal antibody before confirming its cross-reactivity with human and mouse antigens, cross-reactivity with the CD300a antigen, which is known to antagonize the CD300c antigen and has a similar protein sequence to the anti-CD300c monoclonal antibody, was confirmed. More specifically, after treatment with CD300a antigen at concentrations of 0.039 μg / mL, 0.63 μg / mL, and 10 μg / mL, Binding ELISA was performed using the same method as in Example 1.5. The results were... Figure 33 As shown in the image.

[0169] like Figure 33 As shown, it was confirmed that the anti-CD300c monoclonal antibody did not bind to CD300a. Based on the above results, it can be confirmed that it has high binding specificity, to the point that it does not bind to similar sequences of CD300a.

[0170] 6.2. Confirm the ability of anti-CD300c monoclonal antibody to induce differentiation of mouse macrophages (Raw264.7) into M1 cells.

[0171] To confirm whether the anti-CD300c monoclonal antibody could promote the differentiation of mouse macrophages into M1 macrophages, mouse macrophages (Raw264.7) were induced to differentiate into M1 macrophages at a concentration of 1×10⁻⁶ cells / cells. 4 Cells per well were aliquoted into 96-well plates, simultaneously treated with 10 μg / mL anti-CD300c monoclonal antibody, and cultured. TNF-α production was then confirmed using an ELISA kit. Results were... Figure 34 As shown in the image.

[0172] like Figure 34 As shown, the experimental group treated with anti-CD300c monoclonal antibody showed an increase in TNF-α production. These results confirm that it also promotes differentiation into M1 macrophages in both humans and mice.

[0173] 6.3. Inhibitory effect of anti-CD300c monoclonal antibody on the growth of mouse colorectal cancer cells (CT26)

[0174] To confirm whether the anti-CD300c monoclonal antibody also exerted an anti-cancer effect in mice, cell proliferation analysis was performed using CT26 (mouse colon cancer cell line) using the same method as in Example 4.1. The results were... Figure 35 As shown in the image.

[0175] like Figure 35 As shown, the anti-CD300c monoclonal antibody was confirmed to have a cancer treatment effect in mice.

[0176] Example 7: Anti-cancer effect of anti-CD300c monoclonal antibody in vivo

[0177] 7.1. Confirm the inhibitory effect of anti-CD300c monoclonal antibody on cancer growth in vivo.

[0178] To confirm the anti-cancer effect of the anti-CD300c monoclonal antibody in vivo, 2×10⁻⁶ cells were used. 5 A colorectal cancer cell line (CT26) was transplanted subcutaneously into 8-week-old BALB / c mice to establish a syngenic mouse model. All animal breeding and experiments were conducted in a specific pathogen-free (SPF) facility. Figure 36 The experimental method is briefly illustrated in the figure. Twelve days after transplantation of a colorectal cancer cell line, the tumor was introduced into a tumor measuring 50 mm. 3 Up to 100mm 3 Mice were injected with anti-CD300c monoclonal antibody CL7, anti-PD-1 antibody, monoclonal antibody CL7, and anti-PD-1 antibody (Combo), respectively, while the control group received the same amount of phosphate-buffered saline (PBS). More specifically, mice were injected intraperitoneally at a dose of 10 mg / kg twice a week for two weeks (4 times total). Tumor size was then measured after 25 days. The results were... Figure 37 As shown in the image.

[0179] like Figure 37 As shown, it was confirmed that cancer growth was more inhibited in the experimental group that received anti-CD300c monoclonal antibody alone compared with the control group, and cancer growth was more effectively inhibited when used in combination with anti-PD-1 antibody.

[0180] 7.2. Confirm the effect of anti-CD300c monoclonal antibody on increasing tumor-infiltrating lymphocytes in the in vivo tumor microenvironment.

[0181] To confirm the effect of the anti-CD300c monoclonal antibody on tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment (TME), mice euthanized on day 25 of the experiment, which was conducted using the same method as in Example 7.1, were perfused with 1% para-formaldehyde (PFA) to obtain cancer tissue. The obtained cancer tissue was then fixed with 1% PFA and dehydrated sequentially with 10%, 20%, and 30% sucrose solutions. The dehydrated cancer tissue was frozen in an optimal cutting temperature compound and then cut into 50 μm thicknesses using a cryostat. The tumor-infiltrating lymphocyte marker CD8 was then analyzed. + T cells and CD31 + Cancerous blood vessel cells were stained. The results were... Figure 38 As shown in the image.

[0182] like Figure 38 As shown, CD8 in the experimental group treated with anti-CD300c monoclonal antibody was confirmed. + The number of T cells increased compared to the experimental group that received anti-PD-1 antibody injection alone, which confirms that CD300c monoclonal antibody can increase tumor-infiltrating lymphocytes in the tumor microenvironment, thereby having an anti-cancer effect.

[0183] 7.3. Confirmation of the effect of anti-CD300c monoclonal antibody on increasing M1 macrophage activity in vivo.

[0184] To confirm whether the anti-CD300c monoclonal antibody increased M1 macrophages in cancer tissue in vivo, cancer tissue sections prepared using the same method as in Example 7.2 were stained with the M1 macrophage marker iNOS and the M2 macrophage marker CD206. The results were... Figure 39 As shown in the image.

[0185] like Figure 39 As shown, the experimental group treated with anti-PD-1 antibody showed a partial increase in M1 macrophages compared to the control group, while the experimental group treated with anti-CD300c monoclonal antibody showed a significant increase in M1 macrophages, although almost no M2 macrophages were observed. Furthermore, an increase in M1 macrophages was also confirmed in the experimental group treated with a combination of anti-CD300c monoclonal antibody and anti-PD-1 antibody. These results confirm that, compared to existing immunotherapeutic drugs, anti-CD300c monoclonal antibody is more effective in promoting differentiation into M1 macrophages.

[0186] The above results confirm that the anti-CD300c monoclonal antibody of the present invention can bind to the CD300c antigen with high specificity and exhibits cross-species cross-reactivity in mice and other organisms, thus making it suitable for use in various individuals. Furthermore, both in vitro and in vivo studies have confirmed that the anti-CD300c monoclonal antibody can effectively inhibit the proliferation and metastasis of cancer cells by activating T cells and promoting differentiation into M1 macrophages, thereby acting as an immunotherapeutic agent. Moreover, it has been confirmed that the therapeutic effect can be further enhanced by combination therapy with existing immunotherapeutic drugs, thus making it effective for anticancer immunotherapy of various cancers expressing the CD300c antigen.

[0187] The above description of the present invention is merely illustrative. Those skilled in the art will understand that it can be easily modified into other specific forms without altering the technical concept or essential features of the invention. Therefore, all aspects of the embodiments described above should be understood as exemplary, not restrictive.

[0188] Industrial availability

[0189] The anti-CD300c monoclonal antibody of this invention specifically binds to CD300c expression on the surface of cancer cells. Therefore, it can be used not only to treat any cancer that secretes CD300c antigen, but also exhibits cross-species cross-reactivity, making it suitable for anti-cancer treatment in various individuals. Furthermore, because it exhibits anti-cancer effects by acting as an immunotherapeutic agent, effectively inhibiting cancer proliferation, development, and metastasis, it can be effectively used for immunotherapy of various cancers. SEQUENCE LISTING <110> Shancui Kesi Biotechnology Co., Ltd. <120> Compositions containing anti-CD300c monoclonal antibodies for the prevention or treatment of cancer. <130> P22112950WP <150> KR 10-2019-0147496 <151> 2019-11-18 <150> KR 10-2020-0154076 <151> 2020-11-17 <160> 50 <170> PatentIn version 3.5 <210> 1 <211> 714 <212> DNA <213> Artificial Sequence <220> <223> CK1 <400> 1 gaagtgcagc tgctggaaag tggaggtgga ctggtgcagc ctggcggcag cctgcgcctg 60 agctgtgccg ccagcggatt caccttcagc cgctatgcca tgacctgggt tcgccaagca 120 cctggcaaag gcctggaatg ggtgagcagc atgagcggca ccggcggcac cacctattat 180 gccgatagcg tgaaaggtcg ctttaccatc agccgcgata acagcaaaaa caccctgtat 240 ctgcagatga acagcctgcg cgccgaggac accgcagtct actactgtgc ccgcggcgcc 300 tatggctttg atcattgggg acaaggtact ctggtgaccg tgagcagcag tggaggaggt 360 agcggaggtg gtggatctgg aggtggaggt agtgaaatcg tgctgaccca gagccctggc 420 accctgagcc tgagccctgg cgaacgcgca acactgtcat gccgcgccag ccagagcatc 480 ggcaactatc tgaactggta tcagcagaaa ccaggtcagg ctccacgtct gctgatctat 540 gatgccagca acctggaaac cggcatccct gatcgcttct caggatctgg aagcggtacc 600 gattttaccc tgaccatcag ccgcctggaa cctgaggact ttgccgtgta ttattgtcag 660 cagagtagcg ccatccctta taccttcggt cagggcacta aagtggaaat caaa 714 <210> 2 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> CK1 <400> 2 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 Arg Tyr 20 25 30 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Met Ser Gly Thr Gly Gly Thr 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 Gly Ala Tyr Gly Phe Asp His Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 145 150 155 160 Gly Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Asp Ala Ser Asn Leu Glu Thr Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Ala 210 215 220 Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 3 <211> 720 <212> DNA <213> Artificial Sequence <220> <223> CK2 <400> 3 gcggccgaag tgcagctgct ggaaagtgga ggtggactgg tgcagcctgg cggcagcctg 60 cgcctgagct gtgccgccag cggattcacc ttcagcagct atggcatgca ttgggttcgc 120 caagcacctg gcaaaggcct ggaatgggtg agcgccatca gcggcagcgg caccagcatc 180 tattatgccg atagcgtgaa aggccgcttt accatcagcc gcgataacag caaaaacacc 240 ctgtatctgc agatgaacag cctgcgcgcc gaggacaccg cagtctacta ctgtgcccgc 300 ggcggcaccg cctttgatta ttggggacaa ggtactctgg tgaccgtgag cagcggtgga 360 ggaggtagcg gaggtggtgg atctggaggt ggaggtagtg aaatcgtgct gacccagagc 420 cctggcaccc tgagcctgag cctggcgaa cgcgcaacac tgtcatgccg cgccagccag 480 agatcagaca actatctggc ctggtatcag cagaaaccag gtcaggctcc acgtctgctg 540 atctatgatg ccagcaaccg cgccaccggc atccctgatc gcttctcagg atctggaagc 600 ggtaccgatt ttaccctgac catcagccgc ctggaacctg aggactttgc cgtgtattat 660 tgtcagcaga gctatagcac cccttttacc ttcggtcagg gcactaaagt ggaaaccaaa 720 <210> 4 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> CK2 <400> 4 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 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Thr Ser Ile 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 Gly Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Ser 145 150 155 160 Asp Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 They have Tyr Asp with Ser Asn Arg and Thr Gly with Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Ser 210 215 220 Thr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Thr Lys 225 230 235 <210> 5 <211> 723 <212> DNA <213> Artificial Sequence <220> <223> CK3 <400> 5 ggccgagtgc agctgctgga aagtggaggt ggactgtgc agcctggcgg cagcctgcgc 60 ctgagctgtg ccgccagcgg attcaccttc agcagctatg ccatcagctg ggttcgccaa 120 gcacctggca aaggcctgga atgggtgagc gccaccagcg gcagcggccg cgccacctat 180 tatgccgata gcgtgaaagg ccgctttacc atcagccgcg father aaacaccctg tatctgcaga tgaacagcct gcgcgccgag gacaccgcag tctactactg tgcgcgcgat acctggtggg aaggctattt tgatctgtgg ggacaaggta ctctggtgac cgtgagcagc 360 agtggaggag gtagcggagg tggtggatct ggaggtggag gtagtgaaat cgtgctgacc 420. cagagccctg gcaccctgag cctgagccct ggcgaacgcg caacactgtc atgccaggcc 480 agccatatca gcacccatct gaactggtat cagcagaaac caggtcaggc tccacgtctg ctgatctatg gcgccagcag ccgcgccacc ggcatccctg atcgcttctc aggatctgga agcggtaccg attttaccct gaccatcagc cgcctggaac ctgaggactt tgccgtgtat 660 tattgtcagc agtatacac ctatcctcct accttcggtc agggcactaa agtggaaatc aaa 723 <210> 6 <211> 240 <212> PRT <213> Artificial Sequence <220> <223> CK3 <400> 6 Arg 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 Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Thr Ser Gly Ser Gly Arg Ala 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 Asp Thr Trp Trp Glu Gly Tyr Phe Asp Leu Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ser Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr 130 135 140 Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Gln Ala Ser 145 150 155 160 His Ile Ser Thr His Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala 165 170 175 Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro 180 185 190 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 195 200 205 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr 210 215 220 Asn Thr Tyr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 240 <210> 7 <211> 705 <212> DNA <213> Artificial Sequence <220> <223> CL4 <400> 7 cgagtgcagc tgctggaag tggaggtgga ctggtgcagc ctggcggcag cctgcgcctg 60 agctgtgccg ccagcggatt caccttcggc agcaactata tgagctgggt tcgccaagca 120 cctggcaaag gcctggaatg ggtgagcacc atcagcggca gcggcaccag cacctattat 180 gccgatagct tgaaggccg ctttaccatc agccgcgata acaccaaaa caccctgtat 240 ctgcagatga acagcctgcg cgccgaggac accgcagctct actactgtgc ccgcggcatg 300 tggggcatgg atgtgtgggg acaaggtact ctggtgaccg tgagcagcgg tggaggaggt 360 agcggaggtg gtggatctgg aggtggaggt agtcagagcg tgctgaccca gcctcctagc 420 gcctccggta caccaggaca gcgcgtgact attagctgta ccggcaaaca tcggcacacc 480 gtgaactggt accagctact gcctggaact gcacctaagc tgctgatcta tctggatagc 540 gaacgcccta gcggcgtacc tgatcgcttt agcggtagca aatcaggcac cagcgccagc 600 ctggccatca gcggccttcg ctccgaagat gaagccgatt attattgtca gagctatgat 660 agcagcagcg tggtgtttgg tggcggtacc aagctgaccg tgctg 705 <210> 8 <211> 235 <212> PRT <213> Artificial Sequence <220> <223> CL4 <400> 8 Arg 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 Gly Ser Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Gly Ser Gly Thr Ser Thr Tyr Tyr Ala Asp Ser Leu 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 Met Trp Gly Met Asp Val Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr 130 135 140 Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Lys His Arg His Thr 145 150 155 160 Val Asn Trp Tyr Gln Leu Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 165 170 175 Tyr Leu Asp Ser Glu Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 180 185 190 Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser 195 200 205 Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser Ser Val 210 215 220 Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 <210> 9 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> CL5 <400> 9 cgagtgcagc tgctggaaag tggaggtgga ctggtgcagc ctggcggcag cctgcgcctg 60 agctgtgccg ccagcggatt caccttcagc agctatgcca tgcattgggt tcgccaagca 120 cctggcaaag gcctggaatg ggtgagcagc atcagcggcg gcggctatgg cacctattat 180 gccgatagcg tgaaaggccg ctttaccatc agccgcgata acagcaaaaa caccctgtat 240 ctgcagatga acagcctgcg cgccgaggac accgcagtct actactgtgc ccgcagcacc 300 gtgtggggcct ttgatatctg gggacaaggt actctggtga ccgtgagcag cggtggagga 360 ggtagcggag gtggtggatc tggaggtgga ggtagtcaga gcgtgctgac ccagcctcct 420 agcgcctccg gtacaccagg acagcgcgtg actattagct gtagcggcaa caacatcggc 480 agcaaaagcg tgcattgta ccagcaacctg cctggactg cacctaagct gctgatctat 540 gatgtgagca aacgccctag cgagcgtcct gatcgcttta gcggtagcaa atcaggcacc 600 agcgccagtc tggccatcag cgaccttcgc tccgagatg aagccgatta ttattgtcag 660 agctttgata gcagcggcac ctggatcttt ggtggcggta ccaagctgac cgtgctg 717 <210> 10 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> CL5 <400> 10 Arg Val Gln Leu Leu Glu Ser 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 Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Gly Gly Gly Tyr Gly Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Arg Asp Asn Serves 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 Ser Thr Val Trp Ala Phe Asp Ile Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly 130 135 140 Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Asn Asn Ile Gly 145 150 155 160 Ser Lys Ser Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys 165 170 175 Leu Leu Ile Tyr Asp Val Ser Lys Arg Pro Ser Glu Arg Pro Asp Arg 180 185 190 Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Asp 195 200 205 Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Phe Asp Ser 210 215 220 Ser Gly Thr Trp Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 <210> 11 <211> 729 <212> DNA <213> Artificial Sequence <220> <223> CL6 <400> 11 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctacggta tgcattgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc agtcagtggt 300 gcaggtcgtg gtttcttcga ctactgggga caaggtactc tggtcactgt ctcctcaggt 360 ggaggcggtt caggcggagg tggatctggc ggtggcggat cccagtctgt gctgactcag 420 ccaccttcag catctggtac tccaggtcag cgcgtcacca tcagctgcag cggtagcagc 480 agcaacattg gtagcaacta cgtgtactgg tatcagcaac tcccaggcac cgctcctaag 540 ctcctgattt acgaggacaa caagcgtcct agtggtgtgc ctgatcgctt ttctgggtcc 600 aagtctggca cctcagcctc tctggctatc agtggacttc gctccgagga cgaggctgac 660 tattactgca gcagctacac tagcagcagc actgtgatct tcggcggtgg gaccaaactg 720 accgtccta 729 <210> 12 <211> 243 <212> PRT <213> Artificial Sequence <220> <223> CL6 <400> 12 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 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser 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 Val Ser Gly Ala Gly Arg Gly Phe Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala 130 135 140 Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser 145 150 155 160 Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly 165 170 175 Thr Ala Pro Lys Leu Leu Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly 180 185 190 Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu 195 200 205 Ala Ile Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser 210 215 220 Ser Tyr Thr Ser Ser Ser Thr Val Ile Phe Gly Gly Gly Thr Lys Leu 225 230 235 240 Thr Val Leu <210> 13 <211> 720 <212> DNA <213> Artificial Sequence <220> <223> CL7 <400> 13 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc cgctacgcaa tgagctgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgtagcagc 300 cagggtatct tcgacatctg gggacaaggt actctggtca ctgtctcctc aggtggaggc 360 ggttcaggcg gaggtggatc tggcggtggc ggatcccagt ctgtgctgac tcagccacct 420 tcagcatctg gtactccagg tcagcgcgtc accatcagct gcagtggtaa caatatcggt 480 actagacgcg tgcattggta tcagcaactc ccagacaccg ctcctaagct cctgatttac 540 agtaagaaca accgtcctag tggtgtgcct gatcgctttt ctgggtccaa gtctggcacc 600 tcagcctctc tggctatcag tggacttcgc tccgaggacg aggctgacta ttactgcgca 660 gcatgggacg acagcctgag cggtcctgtg ttcggcggtg ggaccaaact gaccgtccta 720 <210> 14 <211> 240 <212> PRT <213> Artificial Sequence <220> <223> CL7 <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 Arg Tyr 20 25 30 Ala Met 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 Ser 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 Ser Ser Gln Gly Ile Phe Asp Ile Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly 130 135 140 Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Asn Asn Ile Gly 145 150 155 160 Thr Arg Arg Val His Trp Tyr Gln Gln Leu Pro Asp Thr Ala Pro Lys 165 170 175 Leu Leu Ile Tyr Ser Lys Asn Asn Arg Pro Ser Gly Val Pro Asp Arg 180 185 190 Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly 195 200 205 Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp 210 215 220 Ser Leu Ser Gly Pro Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 240 <210> 15 <211> 726 <212> DNA <213> Artificial Sequence <220> <223> CL8 <400> 15 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctacgcaa tgagctgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcaaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgtagcggt 300 cgttacgcag acttgacatc tgggggacaa ggtactctgg tcactgtctc ctcaggtgga 360 ggcggttcag gcggaggtgg atctggcggt ggcggatccc agtctgtgct gactcagcca 420 ccttcagcat ctggtactcc aggtcagcgc gtcaccatca gctgcagcgg tagcaacagc 480 aacatcggta acaactacgt gagctggtat cagcaactcc cagacacccc tcctaagctc 540 ctgatttacg acaacaacaa gcgtcctagt ggtgtgcctg atcgcttttc tgggtccaag 600 tctggcacct cagcctctct ggctatcagt ggacttcgct ccgaggacga ggctgactat 660 tactgcagca gctacactag cagcagcact gtgatgttcg gcggtgggac caaactgacc 720 gtccta 726 <210> 16 <211> 242 <212> PRT <213> Artificial Sequence <220> <223> CL8 <400> 16 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 Met 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 Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asn 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 Ser Gly Arg Tyr Ala Asp Leu Thr Ser Gly Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 115 120 125 Gly Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser 130 135 140 Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Asn Ser 145 150 155 160 Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr Gln Gln Leu Pro Asp Thr 165 170 175 Pro Pro Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val 180 185 190 Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala 195 200 205 Ile Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser 210 215 220 Tyr Thr Ser Ser Ser Thr Val Met Phe Gly Gly Gly Thr Lys Leu Thr 225 230 235 240 Val Leu <210> 17 <211> 723 <212> DNA <213> Artificial Sequence <220> <223> CL9 <400> 17 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctactact ggagctgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgtatcgac 300 gtgtacggtt tcgacatctg gggacaaggt actctggtca ctgtctcctc aggtggaggc 360 ggttcaggcg gaggtggatc tggcggtggc ggatcccagt ctgtgctgac tcagccacct 420 tcagcatctg gtactccagg tcagcgcgtc accatcagct gcagcggtag cactagcaac 480 atcggtacta actacgtgta ctggtatcag caactcccag gcaccgctcc taagctcctg 540 atttacgaca acaacaaccg tcctagtggt gtgcctgatc gcttttctgg gtccaagtct 600 ggcacctcag cctctctggc tatcagtgga cttcgctccg aggacgaggc tgactattac 660 tgccagactt gggacagcag cactgacgta gtgttcggcg gtgggaccaa actgaccgtc 720 cta 723 <210> 18 <211> 241 <212> PRT <213> Artificial Sequence <220> <223> CL9 <400> 18 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 Tyr Trp 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 Ser 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 Ile Asp Val Tyr Gly Phe Asp Ile Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly 130 135 140 Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn 145 150 155 160 Ile Gly Thr Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala 165 170 175 Pro Lys Leu Leu Ile Tyr Asp Asn Asn Asn Arg Pro Ser Gly Val Pro 180 185 190 Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile 195 200 205 Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Thr Trp 210 215 220 Asp Ser Ser Thr Asp Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val 225 230 235 240 Leu <210> 19 <211> 720 <212> DNA <213> Artificial Sequence <220> <223> CL10 <400> 19 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctacggta tgcattgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc aagcggttac 300 ggtctgatgg acgtgtgggg acaaggtact ctggtcactg tctcctcagg tggaggcggt 360 tcaggcggag gtggatctgg cggtggcgga tcccagtctg tgctgactca gccaccttca 420 gcatctggta ctccaggtca gcgcgtcacc atcagctgca ctcgtagcag cggtatcatc 480 gcaagcaact acgtgcagtg gtatcagcaa ctcccaggca ccgctcctaa gctcctgatt 540 taccgcaaca accagcgccc tagtggtgtg cctgatcgct tttctgggtc caagtctggc 600 acctcagcct ctctggctat cagtggactt cgctccgagg acgaggctga ctattactgc 660 agcagctacg caggtaacaa caacctggtg ttcggcggtg ggaccaaact gaccgtccta 720 <210> 20 <211> 240 <212> PRT <213> Artificial Sequence <220> <223> CL10 <400> 20 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 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser 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 Ser Gly Tyr Gly Leu Met Asp Val Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr 130 135 140 Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ile Ile 145 150 155 160 Ala Ser Asn Tyr Val Gln Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro 165 170 175 Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp 180 185 190 Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser 195 200 205 Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala 210 215 220 Gly Asn Asn Asn Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 240 <210> 21 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> SK11 <400> 21 ggccgagtgc agctgctgga aagtggaggt ggactggtgc agcctggcgg cagcctgcgc 60 ctgagctgtg ccgccagcgg attcaccttc agcacctatg gcatgcattg ggttcgccaa 120 gcacctggca aaggcctgga atgggtgagc gccatcagcg gcagcggcgg cagcacctat 180 tatgccgata gcgtgaaagg ccgctttacc atcagccgcg ataacagcaa aaacaccctg 240 tatctgcaga tgaacagcct gcgcgccgag gacaccgcag tctactactg tgcccgcggc 300 ctgagcggcc ttgattattg gggacaaggt actctggtga ccgtgagcag cagtggagga 360 ggtagcggag gtagtggatc tggaggtgga ggtagtgaaa tcgtgctgac ccagagccct 420 ggcaccctga gcctgagccc tggcgaacgc gcaacactgt catgccgctc cagccagggc 480 atcaccaact atctggcctg gtatcagcag aaaccaggtc aggctccacg tctgctgatc 540 tatgatgcca gcaaccgcgc caccggcatc cctgatcgct tctcaggatc tggaagcggt 600 accgatttta ccctgaccat cagccgcctg gaacctgagg actttgccgt gtattattgt 660 cagcagagct atagcacccc tctgaccttc ggtcagggca ctaaagtgga aatcaaa 717 <210> 22 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK11 <400> 22 Arg 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 Thr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser 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 Gly Leu Ser Gly Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ser Gly Gly Gly Ser Gly Gly Ser Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ser Ser Gln Gly Ile 145 150 155 160 Thr Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Ser 210 215 220 Thr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 23 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> SK12 <400> 23 ggccgagtgc agctgctgga aagtggaggt ggactggtgc agcctggcgg cagcctgcgc 60 ctgagctgtg ccgccagcgg attcaccttc agcagctatg ccatgcattg ggttcgccaa 120 gcacctggca aaggcctgga atgggtgagc gccatcagcg gcagcggcgg cgatacctat 180 catgccgata gcgtgaaagg ccgctttacc atcagccgcg ataacagcaa aaacaccctg 240 tatctgcaga tgaacagcct gcgcgccgag gacaccgcag tctactactg tacccgcggc 300 ctgagcggct ttgattattg gggacaaggt actctggtga ccgtgagcag cggtggagga 360 ggtagcggag gtggtggatc tggaggtgga ggtagtgaaa tcgtgctgac ccagagccct 420 ggcaccctga gcctgagccc tggcgaacgc gcaacactgt catgccgcgc cagccagagc 480 atcagcagct atctgaacg gtatcagcag aaaccaggtc aggctccacg tctgctgatc 540 tatgatgcca gcaccgcgc ccctggcatc cctgatcgct tctcaggtc tggaagcggt 600 accgatttta ccctgaccat cagccgcctg gaacctgagg acttgccgt gtattattgt 660 cagcagagct atagcatccc tatcaccttc ggtcagggca ctaaagtgga aatcaa 717 <210> 24 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK12 <400> 24 Arg Val Gln Leu Leu Glu Ser 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 Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Asp Thr Tyr His Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Arg Asp Asn Serves 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 Thr Arg Gly Leu Ser Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 145 150 155 160 Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Pro Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Ser 210 215 220 Ile Pro Ile Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 25 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> SK13 <400> 25 ggccgagtgc agctgctgga aagtggaggt ggactggtgc agcctggcgg cagcctgcgc 60 ctgagctgtg ccgccagcgg attcaccttc agcgattatg ccatgagctg ggttcgccaa 120 gcacctggca aaggcctgga atgggtgagc agcatcagca gcagcagcag ctatatctac 180 tataccgata gcgtgaaagg ccgctttacc atcagccgcg ataacagcaa aaacaccctg 240 tatctgcaga tgaacagcct gcgcgccgag gacaccgcag tctactactg tgcccgcggc 300 ggctatggct ttgattattg gggacaaggt accctggtga ccgtgagcag cggtggagga 360 ggtagcggag gtggtggatc tggaggtgga ggtagtgaaa tcgtgctgac ccagagccct 420 ggcaccctga gcctgagccc tggcgaacgc gcaacactgt catgccgcgc cagccagagc 480 atcagcagct atctgaactg gtatcagcag aaaccaggtc aggctccacg tctgctgatc 540 tatagcgcca gcagccgccc acagggcatc cccgatcgct tctcaggatc tggaagcggt 600 accgatttta ccctgaccat cagccgcctg gaacctgagg actttgccgt gtattattgt 660 cagcagtatg atgatctgcc ttttaccttc ggtcagggca ctaaagtgga aatcaaa 717 <210> 26 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK13 <400> 26 Arg 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 Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Ser Ser Ser Ser Tyr Ile Tyr Tyr Thr 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 Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 145 150 155 160 Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Ser Ala Ser Ser Arg Pro Gln Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Asp 210 215 220 Leu Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 27 <211> 714 <212> DNA <213> Artificial Sequence <220> <223> SK14 <400> 27 gaagtgcagc tgctggaaag tggaggtgga ctggtgcagc ctggcggcag cctgcgcctg 60 agctgtgccg ccagcggatt caccttcagc aactttgcga tcgcctgggt tcgccaagca 120 cctggcaaag gcctggaatg ggtgagcgcc atcagcggcc gcggcaccag cacctattat 180 gccgatagcg tgaaaggccg ctttaccatc agccgcgata acagcaaaaa caccctgtat 240 ctgcagatga acagcctgcg cgccgaggac accgcagtct actactgtgc ccgcggcgtg 300 agcggctttg atagctgggg acaaggtact ctggtgaccg tgagcagcgg tggaggaggt 360 agcggaggtg gtggatctgg aggtggaggt agtgaaatcg tgctgaccca gagccctggc 420 accctgagcc tgagccctgg cgaacgcgca acactgtcat gccgcgccag ccagagcatc 480 agcagccatc tggcctggta tcagcagaaa ccaggtcagg ctccacgtct gctgatctat 540 gataccagca accgcgccac cggcatccct gatcgcttct caggatctgg gagcggtacc 600 gattttaccc tgaccatcag ccgcctggaa cctgaggact ttgccgtgta ctattgtcag 660 cagagctata gcaccccttt taccttcggt cagggcacta aagtggaaat caaa 714 <210> 28 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK14 <400> 28 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 Asn Phe 20 25 30 Ala Ile Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Arg Gly Thr Ser 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 Gly Val Ser Gly Phe Asp Ser Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 145 150 155 160 Ser Ser His Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Asp Thr Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Ser 210 215 220 Thr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 29 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> SK15 <400> 29 ggccgagtgc agctgctgga aagtggaggt ggactggtgc agcctggcgg cagcctgcgc 60 ctgagctgtg ccgccagcgg attcaccttc agcagctatg ccatgcattg ggttcgccaa 120 gcacctggca aaggcctgga atgggtgagc gccatcaacg gcagcggcgg cagcacctat 180 tatgccgata gcgtgaaagg ccgctttacc atcagccgcg ataacagcaa aaacaccctg 240 tatctgcaga cgaacagcct gcgcgccgag gacaccgcag tctactactg tgcccgcggc 300 ctgcagggct ttgattattg gggacaaggt actctggtga ccgtgagcag cagtggagga 360 ggtagcggag gtggtggatc tggaggtgga ggtagtgaaa tcgtgctgac ccagagccct 420 ggcaccctga gcctgagccc tggcgaacgc gcaacactgt catgccaggc cagccaggat 480 atcaccaact atctgaactg gtatcagcag aaaccaggtc aggctccacg tctgctgatc 540 tatgatgcca gcagcctgga aaccggcatc cctgatcgtt tctcaggatc tggaagcggt 600 accgatttta ccctgaccat cagccgcctg gaacctgagg actttgccgt gtattattgt 660 cagcagagct atagcacccc tatcaccttc ggtcagggca ctaaagtgga aatcaaa 717 <210> 30 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK15 <400> 30 Arg 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 Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Asn Gly Ser Gly Gly Ser 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 Thr Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Leu Gln Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Gln Ala Ser Gln Asp Ile 145 150 155 160 Thr Tyr Asn Leads Tyr Tyr Lys Pro Gly Gln Ala Pro Arg 165 170 175 The Tyr-Asp-Isolated Ser-Glu-Thr Gly-Ile Pro-Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Ser 210 215 220 Thr Pro Ile Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 31 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> SK16 <400> 31 ggccgagtgc agctgctgga aagtggaggt ggactgtgc agcctggcgg cagcctgcgc 60 ctgagctgtg ccgccagcgg attcaccttc agcagctatg ccatgagctg ggttcgccaa 120 gcacctggca aaggcctgga atgggtgagc gccatcaacg gcagcggcgg cagcaccctg 180 tatgccgata gcgtgaagg ccgctttacc atcagccgcg atacagca aaacaccctg 240 tatctgcaga tgaacagcct gcgcgccgag gacaccgcag tctactactg tgcccgcggc 300 gtgagcggct ttgatagctg gggacaaggt actctggtga ccgtgagcag cggtggagga 360 ggtagcggag gtggtggatc tggaggtgga ggtagtgaaa tcgtgctgac ccagagccct 420 ggcaccctga gcctgagccc tggcgaacgc gcaacactgt catgccgcat cagccagagc 480 atcagcagct atctgaactg gtatcagcag aaaccaggtc aggctccacg tctgctgatc 540 tatgatgcca gcctgcgc caccggcatc cctgatcgct tctcaggatc tggaagcggt 600 accgatttta ccctgaccat cagccgcctg gaacctgagg actttgccgt gtattattgt 660 cagcagagct ataaaacccc tatcaccttc ggtcagggca ctaaagtgga aatcaaa 717 <210> 32 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK16 <400> 32 Arg 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 Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Asn Gly Ser Gly Gly Ser Thr Leu 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 Gly Val Ser Gly Phe Asp Ser Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ile Ser Gln Ser Ile 145 150 155 160 Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Asp Ala Ser Leu Arg Ala Thr Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Lys 210 215 220 Thr Pro Ile Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 33 <211> 714 <212> DNA <213> Artificial Sequence <220> <223> SK17 <400> 33 gaagtgcagc tgctggaaag tggaggtgga ctggtgcagc ctggcggcag cctgcgcctg 60 agctgtgccg ccagcggatt caccttcagc agctattatt ggagctgggt tcgccaagca 120 cctggcaaag gcctggaatg ggtgagcacc atcaccggca gcggcggcag caccgattat 180 gccaacagcg tgaaaggccg ctttaccatc agccgcgata acagcaaaaa caccctgtat 240 ctgcagatga acagcctgcg cgccgaggac accgcagtct actactgtgc caccggcggc 300 ggcatctttg actattgggg acaaggtact ctggtgaccg tgagcagcgg tggaggaggt 360 agcggaggtg gtggatctgg aggtggaggt agtgaaatcg tgctgaccca gagccctggc 420 accctgagcc tgagccctgg cgaacgcgca acactgtcat gccaggccag ccagaccatc 480 agcaactatc tgaactggta tcagcagaaa ccaggtcagg ctccacgtct gctgatctat 540 gatgccagca accgcgccac cggcatccct gatcgcttct caggatctgg aagcggtacc 600 gatttaccc tgaccatcag ccgcctggaa cctgaggact ttgccgtgta ttattgtcag 660 cagtacaaca gctatcctcc tagcttcggt cagggcacta aagtggaaat caaa 714 <210> 34 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> SK17 <400> 34 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 Tyr Trp Expensive Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Thr Gly Ser Gly Gly Ser Thr Asp Tyr Ala Asn 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 Gly Gly Gly Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 130 135 140 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Gln Ala Ser Gln Thr Ile 145 150 155 160 Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 165 170 175 Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg 180 185 190 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 195 200 205 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Ser 210 215 220 Tyr Pro Pro Ser Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 35 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> SL18 <400> 35 cgagtgcagc tgctggaaag tggaggtgga ctggtgcagc ctggcggcag cctgcgcctg 60 agctgtgccg ccagcggatt caccttcagc gattatcata tgcattgggt tcgccaagca 120 cctggcaaag gcctggaatg ggtgagcacc atcagcagca gcggcggcta tacctattat 180 gccgaaagcg tgaaaagccg ctttaccatc agccgcgata acagcaaaaa caccctgtat 240 ctgcagatga acagcctgcg cgccgaggac accgcagtct actactgtgc ccgatcgata 300 cgcctgcctc tggattattg gggacaaggt actctggtga ccgtgagcag cagtggagga 360 ggtagcggag gtggtggatc tggaggtgga ggtagtcaga gcgtgctgac ccagcctcct 420 agcgcctccg gtacaccagg acagcgcgtg actattagct gtagcggcaa caacatcggc 480 agcaaaggcg tgcattgta tcagcactg cctggactg cacctaagct gctgattat 540 gaagatagca aacgccctag cggcgtgcgt gatcgcttta gcggtagcaa atcaggcacc 600 agcgccagcc tggccatcag cggccttcgc tccgagatg aagccgatta ttattgtcag 660 agctatgata gcaccaaagg cgtggtgttt ggtggcggta ccaagctgac cgtgctg 717 <210> 36 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> SL18 <400> 36 Arg Val Gln Leu Leu Glu Ser 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 Asp Tyr 20 25 30 His Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Ser Gly Gly Tyr Thr Tyr Tyr Ala Glu Ser Val 50 55 60 Lys Ser Arg Phe Thr Ile Arg Ser Asp Asn Serves 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 Ser Ile Arg Leu Pro Leu Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly 130 135 140 Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Asn Asn Ile Gly 145 150 155 160 Ser Lys Gly Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys 165 170 175 Leu Leu Ile Tyr Glu Asp Ser Lys Arg Pro Ser Gly Val Arg Asp Arg 180 185 190 Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly 195 200 205 Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser 210 215 220 Thr Lys Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 <210> 37 <211> 720 <212> DNA <213> Artificial Sequence <220> <223> CB301_H3L1_A10 <400> 37 gaggtgcagc tgttggagtc tggtggaggc ttggtacagt ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctacggta tgcattgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgt gcgtggttac 300 ggtgcaatgg acgtgtgggg acaaggtact ctggtcactg tctcctcagg tggaggcggt 360 tcaggcggag gtggatctgg cggtggcgga tcccagtctg tgctgactca gccaccttca 420 gcatctggta ctccaggtca gcgcgtcacc atcagctgca ctcgtagcag cggtagcatc 480 gcaagcaact acgtgcagtg gtatcagcaa ctcccaggca ccgctcctaa gctcctgatt 540 taccgcaaca accagcgccc tagtggtgtg cctgatcgct tttctgggtc caagtctggc 600 acctcagcct ctctggctat cagtggactt cgctccgagg acgaggctga ctattactgc 660 agcagctaca ctactagcag cactctggtg ttcggcggtg ggaccaaact gaccgtccta 720 <210> 38 <211> 240 <212> PRT <213> Artificial Sequence <220> <223> CB301_H3L1_A10 <400> 38 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Ser 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 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser 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 Val Arg Gly Tyr Gly Ala Met Asp Val Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr 130 135 140 Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ser Ile 145 150 155 160 Ala Ser Asn Tyr Val Gln Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro 165 170 175 Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp 180 185 190 Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser 195 200 205 Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr 210 215 220 Thr Ser Ser Thr Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 240 <210> 39 <211> 723 <212> DNA <213> Artificial Sequence <220> <223> CB301_H3L1_A12 <400> 39 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctacgcaa tgcattgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcaaaggac actgccgtgt attactgcgc aagcggctac 300 ggtctgatgg acgtatgggg acaaggtact ctggtcactg tctcctcagg tggaggcggt 360 tcaggcggag gtggatctgg cggtggcgga tcccagtctg tgctgactca gccaccttca 420 gcatctggta ctccaggtca gcgcgtcacc atcagctgca ctggtactag cagcgacgtg 480 ggtaactaca acctggtgag ctggtatcag caactcccag gcaccgctcc taagctcctg 540 atttacagca acaaccagcg ccctagtggt gtgcctgatc gcttttctgg gtccaagtct 600 ggcacctcag cctctctggc tatcagtgga cttcgctccg aggacgaggc tgactattac 660 tgcagcagct acactggtag caacgctctg ttgttcggcg gtgggaccaa actgaccgtc 720 cta 723 <210> 40 <211> 241 <212> PRT <213> Artificial Sequence <220> <223> CB301_H3L1_A12 <400> 40 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 Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser 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 Lys Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ser Gly Tyr Gly Leu Met Asp Val Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr 130 135 140 Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val 145 150 155 160 Gly Asn Tyr Asn Leu Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala 165 170 175 Pro Lys Leu Leu Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro 180 185 190 Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile 195 200 205 Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr 210 215 220 Thr Gly Ser Asn Ala Leu Leu Phe Gly Gly Gly Thr Lys Leu Thr Val 225 230 235 240 Leu <210> 41 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> CB301_H3L1_E6 <400> 41 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc agctacgcaa tgagctgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgctggcat 300 tacagcttcg actactgggg acaaggtact ctggtcactg tctcctcagg tggaggcggt 360 tcaggcggag gtggatctgg cggtggcgga tcccagtctg tgctgactca gccaccttca 420 gcatctggta ctccaggtca gcgcgtcacc atcagctgcc gtggtaacaa catcggtagc 480 aagcgtgtgc attggtatca gcaactccca ggcaccgctc ctaagctcct gatttacagc 540 tacaaccacc gtcctagcgg tgtgcctgat cgcttttctg ggtccaagtc tggcacctca 600 gcctctctgg ctatcactgg acttcgctcc gaggacgaag ctgactatta ctgcaacact 660 tgggacgaca gcctggaggg tcctgtgttc ggcggtggga ccaaactgac cgtccta 717 <210> 42 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> CB301_H3L1_E6 <400> 42 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 Met 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 Ser 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 Trp His Tyr Ser Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr 130 135 140 Pro Gly Gln Arg Val Thr Ile Ser Cys Arg Gly Asn Asn Ile Gly Ser 145 150 155 160 Lys Arg Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 165 170 175 Leu Ile Tyr Ser Tyr Asn His Arg Pro Ser Gly Val Pro Asp Arg Phe 180 185 190 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 195 200 205 Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Thr Trp Asp Asp Ser 210 215 220 Leu Glu Gly Pro Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 <210> 43 <211> 714 <212> DNA <213> Artificial Sequence <220> <223> CB301_H3L1_F4 <400> 43 gaggtgcagc tgttggagtc tggtggaggc ttggtacagc ctggaggttc tcttcgcctc 60 tcctgtgcag cctccggatt cactttcagc ggctacgcaa tgagctgggt cagacaggca 120 ccaggtaagg gactggagtg ggtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgtagtcct 300 agcggtctgt tcgactactg gggacaaggt actctggtca ctgtctcctc aggtggaggc 360 ggttcaggcg gaggtggatc tggcggtggc ggattccagt ctgtgctgac tcagccacct 420 tcagcatctg gtactccagg tcagcgcgtc accatcagct gcggtggtaa caacatcggt 480 agcaagcgtg tgcattggta tcagcaactc ccaggcaccg ctcctaagct cctgatttac 540 aacactagca acaagcatag cggtgtgcct gatcgctttt ctgggtccaa gtctggcacc 600 tcagcctctc tggctatcag tggacttcgc tccgaggacg aggctgacta ttactgcagc 660 agctacctac agcagcactc tctgttcggc ggtgggacca aactaaccgt ccta 714 <210> 44 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> CB301_H3L1_F4 <400> 44 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 Gly Tyr 20 25 30 Ala Met 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 Ser 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 Ser Pro Ser Gly Leu Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Phe Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly 130 135 140 Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Gly Gly Asn Asn Ile Gly 145 150 155 160 Serving Lys Arg Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys 165 170 175 Leu Leu Ile Tyr Asn Thr Ser Asn Lys His Ser Gly Val Pro Asp Arg 180 185 190 Phe To Be Gly To Be Lys To Be Gly To Be Thr To Be Ala To Be Leu To Be Ile To Be Gly 195 200 205 Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Leu Gln 210 215 220 Gln His Ser Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235 <210> 45 <211> 729 <212> DNA <213> Artificial Sequence <220> <223> CB301_H3L1_G11 <400> 45 gaggtgcagc tgttggagtc tggtggaggc tggtacagc ctggaggttc ctcttccgcc 60 tcctcctgtg cagcctccgg attcacttc agcagctacg caatgagctg ggtcagacag 120 gcaccaggta agggactgga gtgggtctct gcaattagcg gtagcggtgg tagcacttac 180 tacgcagaca gcgtgaaggg tcgctcacc atctcacgcg acaactccaa gaacaccctg 240 tacctgcaga tgaacagcct tcgcgcagag gacactgccg tgtattactg cacacgtttc 300 gtgggtgcaa tcggtgcatt cgactactgg ggacaaggta ctctggtcac tgtctcctca 360 ggtggaggcg gttcaggcgg aggtggatct ggcggtggcg gatcccagtc tgtgctgact 420 cagccacctt cagcatctgg tactccaggt cagcgcgtca ccatcagctg cagtggtaac 480 aacatcggta gccgtagcgt gcattggtat cagcaactcc caggcaccgc tcctaagctc 540 ctgatttacc gcaacaacca gcgccctagt ggtgtgcctg atcgctttc tgggtccaag 600 tctggcacct cagcctctct ggctatcagt ggacttcgct ccgaggacga ggctgactat 660 tactgcgcag catgggacga cagcctgagc ggtcctgtgt tcggcggtgg gaccaaactg 720 accgtccta 729 <210> 46 <211> 243 <212> PRT <213> Artificial Sequence <220> <223> CB301_H3L1_G11 <400> 46 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Ser Ser Ala Ser Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 20 25 30 Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Thr Arg Phe Val Gly Ala Ile Gly Ala Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser 130 135 140 Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Asn 145 150 155 160 Asn Ile Gly Ser Arg Ser Val His Trp Tyr Gln Gln Leu Pro Gly Thr 165 170 175 Ala Pro Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val 180 185 190 Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala 195 200 205 Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala 210 215 220 Trp Asp Asp Ser Leu Ser Gly Pro Val Phe Gly Gly Gly Thr Lys Leu 225 230 235 240 Thr Val Leu <210> 47 <211> 738 <212> DNA <213> Artificial Sequence <220> <223> CB301_OPALTL_B5 <400> 47 gaggtgcagc tgttggagtc tggtggaggc tggtacagc ctggaggttc tctcgccctc 60 tcctgtgcag cctccggatt cacttcagc cattacgcaa tgagctggt cagacaggca 120 ccaggtagg gactggagtg gtctctgca attagcggta gcggtggtag cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgtggttgg 300 gacagcccta ctctgacata cttcgacagc tggggacaag gtactctggt cactgtctcc 360 tcaggtggag gcggttcagg cggaggtgga tctggcggtg gcggatccca gtctgtgctg 420 actcagccac cttcagcatc tggtactcca ggtcagcgcg tcaccatcag ctgcagcggt 480 actagcagca acatcggtaa caacgacgtg agctggtatc agcaactccc aggcaccgct 540 cctaagctcc tgatttacca ggacactaag cgtcctagcg gtgtgcctga tcgcttttct 600 gggtccaagt ctggcacctc agcctctctg gctatcagtg gacttcgctc cgaggacgag 660 gctgactatt actgcgcagc atgggacgac agcctgagcg gtcctgtgtt cggcggtggg 720 accaaactga ccgtccta 738 <210> 48 <211> 246 <212> PRT <213> Artificial Sequence <220> <223> CB301_OPALTL_B5 <400> 48 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 Ala Met 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 Ser 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 Gly Trp Asp Ser Pro Thr Leu Thr Tyr Phe Asp Ser Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro 130 135 140 Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 145 150 155 160 Thr Ser Ser Asn Ile Gly Asn Asn Asp Val Ser Trp Tyr Gln Gln Leu 165 170 175 Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Gln Asp Thr Lys Arg Pro 180 185 190 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala 195 200 205 Ser Leu Ala Ile Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr 210 215 220 Cys Ala Ala Trp Asp Asp Ser Leu Ser Gly Pro Val Phe Gly Gly Gly 225 230 235 240 Thr Lys Leu Thr Val Leu 245 <210> 49 <211> 717 <212> DNA <213> Artificial Sequence <220> <223> CB301_OPALTL_E6 <400> 49 gaggtgcagc tgttggagtc tggtggaggc tggtacagc ctggaggttc tctcgccctc 60 tcctgtgcag cctccggatt cacttcagc agctacggta tgcattggt cagacaggca 120 ccaggtagg gactggagtg gtctctgca atcagcggta gcggtggtta cacttactac 180 gcagacagcg tgaagggtcg cttcaccatc tcacgcgaca actccaagaa caccctgtac 240 ctgcagatga acagccttcg cgcagaggac actgccgtgt attactgcgc acgctggcat 300 tacagcttcg actactgggg acaaggtact ctggtcactg tctcctcagg tggaggcggt 360 tcaggcggag gtggatctgg cggtggcgga tcccagtctg tgctgactca gccaccttca 420 gcatctggta ctccaggtca gcgcgtcacc atcagctgca gcggtagcag cagcaacatc 480 ggtaacaact acgtgagctg gtatcagcaa ctcccaggca ccgctcctaa gctcctgatt 540 taccgcaaca accagcgccc tagtggtgtg cctgatcgct tttctgggtc caagtctggc 600 acctcagcct ctctggctat cagtggactt cgctccgagg acgaggctga ctattactgc 660 cagagctacg acaacagcaa cgtgctgttc ggcggtggga caaactgac cgtccta 717 <210> 50 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> CB301_OPALTL_E6 <400> 50 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 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Tyr 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 Trp His Tyr Ser Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr 130 135 140 Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile 145 150 155 160 Gly Asn Asn Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro 165 170 175 Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp 180 185 190 Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser 195 200 205 Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp 210 215 220 Asn Ser Asn Val Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 225 230 235

Claims

1. An anti-CD300c monoclonal antibody, characterized in that, The amino acid sequence of the anti-CD300c monoclonal antibody is sequence 14.

2. The anti-CD300c monoclonal antibody according to claim 1, characterized in that, The aforementioned anti-CD300c monoclonal antibody exhibits cross-species cross-reactivity.

3. The anti-CD300c monoclonal antibody according to claim 2, characterized in that, The aforementioned cross-reactivity refers to the cross-reactivity between human antigens and mouse antigens.

4. Use of a pharmaceutical composition in the preparation of a medicament for the prevention or treatment of cancer, said pharmaceutical composition comprising an anti-CD300c monoclonal antibody as an active ingredient, wherein, The amino acid sequence of the above antibody is sequence 14; The cancer is a cancer that expresses the CD300c protein on the surface of cancer cells, and the cancer is one or more selected from the group consisting of colorectal cancer, lung cancer, and breast cancer.

5. The use according to claim 4, characterized in that, The above-mentioned pharmaceutical composition also includes an immunomodulatory anticancer drug.

6. The use according to claim 5, characterized in that, The aforementioned immunotherapeutic drugs are selected from one or more of the following groups: anti-PD-1, anti-PD-L1, anti-CTLA-4, anti-KIR, anti-LAG3, anti-CD137, anti-OX40, anti-CD276, anti-CD27, anti-GITR, anti-TIM3, anti-41BB, anti-CD226, anti-CD40, anti-CD70, anti-ICOS, anti-CD40L, anti-BTLA, anti-TCR, and anti-TIGIT.

7. The use according to claim 4, characterized in that, The above-mentioned drug composition inhibits the proliferation, survival, metastasis, recurrence, or drug resistance of cancer.

8. An immunomodulatory anticancer drug, characterized in that, The immunotherapeutic drug contains an anti-CD300c monoclonal antibody as an active ingredient, wherein the amino acid sequence of the antibody is sequence 14.

9. The immunomodulatory anticancer drug according to claim 8, characterized in that, The aforementioned immunotherapeutic drugs inhibit cancer proliferation, survival, metastasis, recurrence, or anticancer drug resistance.

10. The immunomodulatory anticancer drug according to claim 8, characterized in that, The cancers mentioned above are selected from one or more of the group consisting of colorectal cancer, lung cancer, and breast cancer.

11. An adjuvant for anticancer therapy, said adjuvant comprising an anti-CD300c monoclonal antibody as an active ingredient, wherein, The amino acid sequence of the above antibody is sequence 14.

12. The adjuvant according to claim 11, characterized in that, The above-mentioned adjuvants enhance the anti-cancer treatment effect by activating the immune function of immune cells.

13. The adjuvant according to claim 11, characterized in that, The aforementioned anti-cancer treatment is radiation therapy.

14. The adjuvant according to claim 11, characterized in that, The above-mentioned anti-cancer treatments are anti-cancer drug therapies.

15. The adjuvant according to claim 11, characterized in that, The aforementioned anticancer therapy is an immunotherapy using anticancer drugs.

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