Method for preparing and screening cell line expressing bispecific antibody

A bispecific antibody, cell line technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, cells modified by introducing foreign genetic material, etc., can solve the problem of time-consuming, cumbersome identification process, and low accuracy. question

Inactive Publication Date: 2015-08-05
WUHAN YZY BIOPHARMA CO LTD
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  • Abstract
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Problems solved by technology

[0005] Therefore, in order to solve the shortcomings of the current method for screening the stability of cell lines expressing bispecific antibodies, such as time-consuming, c

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  • Method for preparing and screening cell line expressing bispecific antibody
  • Method for preparing and screening cell line expressing bispecific antibody
  • Method for preparing and screening cell line expressing bispecific antibody

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Embodiment Construction

[0038] 1. Experimental materials

[0039] The bispecific antibody used in this experiment is a monovalent single-chain bispecific antibody (MSBODY (monomer and ScFv bispecific antibody)). MSBODY was prepared by Wuhan Youzhiyou Biopharmaceutical Co., Ltd. according to the method disclosed in PCT / CN2012 / 084982, and can be purchased from this company. MSBODY is composed of two units: one unit is a light chain-heavy chain pair specific to tumor cells or microorganisms, called a monovalent unit; the other unit is a fusion peptide, which contains a single-chain variable fragment ( scFv) and an Fc fragment with a CH2 domain and a CH3 domain, wherein the fusion peptide is specific for immune cells and is called a single-chain unit. Wherein, the light chain constant region (CL) of the monovalent unit is completely consistent with the light chain constant region of human IgG antibody. Monovalent units specifically bind to different tumor target antigens or microbial target antigens, s...

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Abstract

The present invention provides a method for preparing and screening a cell line expressing bispecific antibody. The method comprises: constructing two plasmids expressing bispecific antibody, wherein the two plasmids contain double promoters and respectively express different fluorescent proteins; and carrying out transformation, culture and extraction on the two recombinant plasmids, co-transfecting the two recombinant plasmids into host cells, screening the positive monoclonal cell line expressing the double fluorescence, and evaluating the yield and the stability according to the fluorescence intensity. With the method of the present invention, the high yield cell line can be conveniently and rapidly screened in advance, the test period can be shortened, the manpower and material resource investment can be reduced, and the accuracy is high.

Description

technical field [0001] The invention relates to the field of antibody preparation, in particular to a method for preparing and screening cell lines expressing bispecific antibodies. Background technique [0002] Bispecific antibody (bispecific antibody, hetero conjugate antibody) is a class of antibody molecules with dual functions. The Fab segments in bivalent antibodies have different specificities and can bind to different ligands. Anti-tumor and anti-immune active cell CD16 or CD3 bispecific antibody not only has the effect of activating NK cells or T cells, but also can specifically bind tumor cells through the anti-tumor Fab segment to increase the concentration of local NK cells or T cells , enhance the ability of effector molecules to kill tumors. Immunocytokine can also be prepared by fusing antibodies with certain cytokines to increase the concentration of cytokines near tumor cells, stimulate the immune function of the body, effectively kill tumors, and reduce si...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/13C12N5/10
Inventor 张敬刘晓燕范克索周鹏飞
Owner WUHAN YZY BIOPHARMA CO LTD
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