Preparation method of BSA monoclonal antibody

An antigen and monoclonal technology, applied in chemical instruments and methods, anti-animal/human immunoglobulin, material inspection products, etc., can solve problems such as shortage and death, and achieve the effect of increasing production

Inactive Publication Date: 2021-04-23
TIANJIN AMCELLGENE ENG
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Problems solved by technology

In the HAT medium, unfused myeloma cells lack hypoxanthine-guanine-phosphoribosyltransferase and cannot use the salvage pathway to synthesize DNA and die. Although unfused lymphocytes have hypoxanthine-guanine-phosphate Ribosyltransferase, but itself cannot survive long-term in vitro and gradually dies

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  • Preparation method of BSA monoclonal antibody
  • Preparation method of BSA monoclonal antibody
  • Preparation method of BSA monoclonal antibody

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Experimental program
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Embodiment

[0038]A BSA monoclonal preparation method, the BSA mono-prepared method is:

[0039](1) Preparation polypeptide: First, a polypeptide having a purity of 10 mg of a purity of 90%, wherein the peptide sequence of the polypeptide is KEACFAVEGPKLVVSTQT, and the 5 mg polypeptide is coupled to KLH;

[0040](2) Animal Immunization: Select five small mice, first immunize, the KLH coupling polypeptide is mixed with an equal volume Fuche complete adjuvant, etc. After emulsification, the mice are subcutaneously immunized, and 400 ul is injected under each small mouse skin Emulsion; then carry out a second immunization, 1 mg / ml of KLH antigen is mixed with an equivalent Fuche's incomplete adjuvant to mix and emulsify, and an 200ul emulsion is injected to the abdominal cavity of each small mouse, and repeat The second immunity was subjected to the third immunization of the mice. After 10 days of the third immunity, blood was passed through the eye of each small mice, and centrifugal treatment was ca...

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Abstract

The invention provides a preparation method of a BSA monoclonal antibody. The method comprises the following steps: preparing polypeptide: firstly, preparing 10mg of polypeptide with the purity of 90%, and coupling 5mg of polypeptide with KLH; carrying out animal immunization: selecting five mice, firstly carrying out primary immunization, then carrying out secondary immunization, repeating the secondary immunization step to carry out third immunization on the mice, and then carrying out fourth immunization; carrying out cell fusion; screening: selecting monoclonal holes with high positive values; culturing and amplifying the selected monoclonal hybridoma cells and cryopreserving the cells; and obtaining purified monoclonal antibodies. A large number of monoclonal antibodies can be prepared, the prepared antibodies are stored at minus 20 DEG C after being subpackaged and stored at 2-8 DEG C in a short time, when the antibodies are used, the solution only needs to be diluted according to the needed concentration and prepared for immediate use, the antibody production amount is greatly increased, it is found through inspection that the monoclonal antibodies can be combined with BSA and cannot be combined with HAS, and a foundation is laid for future medical development.

Description

Technical field[0001]The present invention belongs to the field of monocompansion preparation methods, and more particularly to a method of preparing a BSA monocompanion.Background technique[0002]The monoclonal antibody is a height produced by a single B cell clone, only for an antibody of a particular antigenic epitope. Hybridoma techniques are usually prepared, and hybridoma antibody techniques are based on cell fusion technology, which combines allergic B cells with secretion-specific antibody capabilities and myeloma cells with infinite reproductive capabilities into B cell hybridomas.[0003]The procedure of the monoclonal preparation is: 1. Immunogenic animal: Immunogen is immunized by the purpose of antigen, so that mice produce sensitization B lymphocytes, generally selected 6-8 patients with Balb / C mice, according to The pre-developed immunization scheme is immunoaded, antigen enters the peripheral immune organ through blood circulation or lymph cycles, stimulates the corre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18G01N33/68G01N33/577
CPCC07K16/18G01N33/68G01N33/577C07K2317/56G01N2333/765
Inventor 韩之波
Owner TIANJIN AMCELLGENE ENG
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