Monoclonal antibody with liver cancer resisting activity and application thereof

A monoclonal antibody and antibody technology, applied in the fields of application, antibody, anti-tumor drug, etc., can solve the problem of negative regulation of GPC3 expression, and achieve strong binding, high titer, and high specificity

Active Publication Date: 2013-04-10
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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Problems solved by technology

[0007] The role of GPC3 expression in hepatocytes and the signaling pathways involved: When the expression of GPC3 in normal liver cells increases, cell division and proliferation decrease, and blocking the expression of GPC3 promotes the growth of hepatocytes. However, the situation is different in liver cancer cells, and the expression of GPC3 does not play a negative role. sex regulation

Method used

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  • Monoclonal antibody with liver cancer resisting activity and application thereof
  • Monoclonal antibody with liver cancer resisting activity and application thereof
  • Monoclonal antibody with liver cancer resisting activity and application thereof

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Experimental program
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Embodiment 1

[0027] Embodiment 1, preparation of anti-liver cancer active monoclonal antibody

[0028] 1. Main experimental materials

[0029] Main cell lines: mouse myeloma cell line Sp2 / 0, purchased from Nanjing Kaiji Biotechnology Development Co., Ltd., liver cancer cell line HepG2, purchased from the Basic Medical Cell Center of Hebei Medical University, and COS7, purchased from Beijing Hongbokang Pharmaceutical Technology Co., Ltd. ; BALB / c female mice aged 6-8 weeks were provided by the Experimental Animal Center of the Academy of Military Medical Sciences (permit number: scxk-(army)-2007-004).

[0030] Main reagents: cloning vector pMD18-T was purchased from TaKaRa Company, prokaryotic expression vectors pET32a and pGEX-4T-1 were purchased from Novagen and Amersham Company respectively, and pcDNA3.1(+) was a product of Invitrogen. Escherichia coli competent DH5α, BL21(DE3), mini-plasmid extraction kit and DNA gel recovery kit were purchased from Beijing Biomed Biotechnology Company, ...

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Abstract

The invention discloses a monoclonal antibody with a liver cancer resisting activity and application thereof. The monoclonal antibody can perform specific binding with glypican-3 (GPC3) antigen epitope; and the amino acid sequence of the GPC3 antigen epitope is 359th-580th amino acid residue from an amino terminal with Gen Bank accession Number of AAA98132.1. In the invention, a high-valence high-specificity mouse-anti-human GPC3-C monoclonal antibody with an immunological activity and a cancer cell poisoning activity is prepared, thereby establishing an experimental foundation for studying expression and distribution of GPC3 in tissues and cells, deeply studying biological function of GPC3 protein and early detecting liver cancer in clinic and providing a non-toxicity auxiliary medicament.

Description

technical field [0001] The invention relates to an anti-liver cancer active monoclonal antibody and its application. Background technique [0002] Primary liver cancer is one of the most common malignant tumors in the world, and its incidence is on the rise worldwide. my country is the country where the incidence of liver cancer is concentrated in the world. The prognosis of hepatocellular carcinoma (HCC) is extremely poor, and the key to prolonging the survival of patients is early diagnosis and early treatment. Glypican-3 (GPC3) is abnormally expressed in liver cancer cells, but not on the surface of normal liver cells and other non-tumor liver disease cells. And it is missing in ovarian cancer, breast cancer, lung cancer, mesothelioma, kidney cancer and other tumors, and it is a potential specific marker for liver cancer. GPC3 is a cell membrane protein with a molecular weight of 60-70kD. It is a member of the heparan sulfate proteoglycan family and binds to the cell ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N5/20C12N15/12G01N33/577A61K39/395A61P35/00C12R1/91
Inventor 周育森李军锋张慧娜于虹寇志华陈万荣
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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