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Antibody screening method based on single cell chip technology

A chip technology and antibody screening technology, applied in chemical instruments and methods, anti-animal/human immunoglobulin, material inspection products, etc., can solve the problem of inability to achieve large-throughput screening and obtain tens of thousands of antibody sequences and affinities at the same time Data and other issues

Inactive Publication Date: 2017-09-29
GUANGDONG INST OF APPLIED BIOLOGICAL RESOURCES
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AI Technical Summary

Problems solved by technology

At present, there are many technical difficulties in antibody screening and process development in China. For example, most of the antibody screening methods using hybridoma cell technology have many disadvantages. It is impossible to achieve large-throughput screening and obtain tens of thousands of antibody sequences at the same time. and affinity data, etc., resulting in a lower probability of obtaining the optimal antibody sequence

Method used

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Experimental program
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Effect test

Embodiment 1

[0015] A method for screening antibodies based on single-cell chip technology, comprising the following steps:

[0016] Step 1) Immunize mice with cells expressing specific surface antigens, extract mouse spleen cells for ELISA analysis;

[0017] 100 μL of spleen cell suspension (5×106 cells / mL) was used as the antigen, and 10 μL of PEG-precipitated phage antibody was added to 100 μL with 3% BSA / PBS. At the same time, PBS blank, VCSM13 negative control, HRP-goat anti-M13 (with 3 %BSA / PBS (diluted 4000 times) was used as the secondary antibody, and the A value > 2 times the A value of the negative control was used as the criterion for judging positive clones. After color development, observe under an inverted microscope and take pictures.

[0018] step 2)

[0019] According to the results, the single cells are attached to the cell chip and combined with the fluorescently-labeled male-specific antigen (serologically detected H-Y antigens) to obtain positive clone information a...

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Abstract

The invention discloses an antibody screening method based on a single cell chip technology. The method comprises the following steps: 1) expressing a cellular immunized mouse with a special surface antigen and extracting a spleen cell of the mouse for ELISA analysis; 2) attaching the single cell to a cell chip according to the result, and combining the cell chip with fluorescently-labeled antigen to obtain positive clone information and affinity information of antibody; and 3) performing on-site gene amplification on the antibody meeting the standard to obtain an antibody variable region sequence. The method disclosed by the invention can quickly obtain a lot of sequence and affinity data of the antibody. The screening method disclosed by the invention lays a good foundation for pharmaceutical research field.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an antibody screening method based on single cell chip technology. Background technique [0002] In recent years, new antibody drugs have shown good curative effects in the treatment of major diseases such as malignant tumors and autoimmune diseases. The development of antibody drugs at home and abroad has become a new hot spot, and the competition is extremely fierce. In the development process of new antibody drugs, a number of key technologies need to be established and applied, including antibody screening and functional confirmation, cell line construction and process development, pilot scale-up, and preclinical research. At present, there are many technical difficulties in antibody screening and process development in China. For example, most of the antibody screening methods using hybridoma cell technology have many disadvantages. It is impossible to achieve large-throughput...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53C07K16/18
CPCC07K16/18C07K2317/55G01N33/53
Inventor 杨威王笑非毛明汪巨峰王茜莎韩重肖百全郭健敏黄远铿王俐梅
Owner GUANGDONG INST OF APPLIED BIOLOGICAL RESOURCES
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