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Polypeptide chip and preparation process thereof, and method for preparing monoclonal antibody group using the same

A monoclonal antibody and polypeptide chip technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as inability to clone at the same time, and achieve the effect of avoiding continuous repetition, promoting research, and saving time and money.

Inactive Publication Date: 2006-04-26
SHANGHAI BIOCHIP
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Problems solved by technology

In actual work, when the fusion rate is high, a large number of antibody-producing hybridomas are often obtained, and they cannot be cloned at the same time. The positive cells can only be screened according to the OD value read by the microplate reader, and the clones are constructed. This is very likely to cause differences in the results. The monoclonal antibody of the cell line acts on the same epitope

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  • Polypeptide chip and preparation process thereof, and method for preparing monoclonal antibody group using the same
  • Polypeptide chip and preparation process thereof, and method for preparing monoclonal antibody group using the same

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Embodiment Construction

[0030] The preparation method of the polypeptide chip disclosed by the present invention is as follows:

[0031] 1. Design and synthesis of peptides

[0032] On the one hand, according to the antigens of the monoclonal antibody groups with different epitopes prepared as needed, use bioinformatics software to analyze the amino acid sequences of the antigens, predict their possible epitopes, and synthesize corresponding polypeptides; on the other hand, According to the amino acid sequence of the antigen, a series of polypeptides containing repeated sequences of fixed length are synthesized.

[0033] 2. Preparation of Peptide Chip

[0034] (1) Preparation of solid-phase peptide chip

[0035] (a) Dissolve the peptide synthesized in the first step in a certain buffer system, phosphate buffer containing 0.05% BSA is recommended, and the peptide concentration is about 100 μg / ml;

[0036] (b) Transfer the dissolved polypeptide solution into a 96-well plate or a 384-well plate, abou...

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Abstract

The invention discloses a polypeptide chip of monoclonal antibody group screening with different antigen epitopes, which comprises the following steps: 1. synthesizing polypeptide according to the screened antigen of monoclonal antibody group; preparing polypeptide chip; 2. mixing the cell suspension of immune mouse spleen with the antigen and myeloid oncocyte at certain proportion; fusing two cell through fluxing agent to generate hybrid oncocyte; 3. collecting the hybrid oncocyte; cultivating for 7-8 days; making suspending liquid; 4. adapting polypeptide chip of different antigen epitopes to screen and identify the suspending liquid of hybrid oncocyte; 5. establishing a group of monoclonal antibody cell plant of different antigen epitopes with the positive clone screened and identified by polypeptide chip; 6. generating monoclonal antibody group of different antigen epitopes through cultivation enlargement and purity segregation.

Description

technical field [0001] The invention belongs to the field of immunohistochemical biochips of life sciences, and specifically relates to a polypeptide chip and a preparation method thereof, and a method for simultaneously preparing monoclonal antibody groups with different antigen epitopes on a large scale by using the same. Background technique [0002] With the implementation and advancement of the Human Genome Project, life science research has entered the post-genome era. In this era, the main research object of life science is functional genomics, including structural genome research and proteome research. The traditional way of studying a single protein can no longer meet the requirements of the post-genome era. This is because: (1) The occurrence of life phenomena is often influenced by multiple factors, which must involve multiple proteins. (2) The participation of multiple proteins is interwoven into a network, or occurs in parallel, or is a cascade of causality. ...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/531G01N33/546G01N33/547G01N33/551
Inventor 张庆华吴船缪金明宋凯王升年赵芹奚佳捷
Owner SHANGHAI BIOCHIP
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