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Hybridoma cell and preparation method and application thereof

A technology of hybridoma cells and hybridoma cell lines, which is applied in the field of monoclonal antibody hybridoma cell lines and its preparation, can solve the problems of wide application of monoclonal antibodies and unsatisfactory effects of monoclonal antibodies, so as to avoid the spread of the epidemic and Effects of social panic, shortened diagnosis time, and improved quality and level

Inactive Publication Date: 2011-10-05
云南省地方病防治所
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AI Technical Summary

Problems solved by technology

At present, the effect of monoclonal antibody secretion by hybridoma strains obtained by applying cell hybridoma technology in the field of plague research is not ideal. Therefore, monoclonal antibodies have not been widely used in the diagnosis of plague.

Method used

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  • Hybridoma cell and preparation method and application thereof
  • Hybridoma cell and preparation method and application thereof
  • Hybridoma cell and preparation method and application thereof

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Experimental program
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Embodiment Construction

[0017] Specific embodiments of the present invention will be described in detail below.

[0018] The main reagents used in the present invention are as follows:

[0019] Yersinia pestis F1 antigen (self-made), RPMI-1640 medium (Gibco company), sodium pyruvate (Sigma company), L-glutamine (USB company), Hepes (USB company), polyethylene glycol ( PEG) (Sigma Company), HAT freeze-dried powder and HT freeze-dried powder (Sigma Company), 8-azaguanine (8-AG) (Sigma Company), dimethylsulfoxide (DMSO) (Sigma Company), Rabbit anti-mouse IgG (enzyme-labeled antibody) (Sigma Company), enhanced calf serum (FBCS) (Hyclone Company), Freund's complete and incomplete adjuvant (Sigma Company), tetramethylbenzidine (TMB) (Shanghai Livzon Dongfeng Biotechnology Company).

[0020] Main instruments used in the present invention are as follows:

[0021] CO 2 Incubator (Heraeus, Germany), inverted microscope (Nikon, Japan), sterilizing filter (AG.37075, Germany) and 0.20 μm filter membrane (Germ...

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Abstract

The invention relates to a method for preparing a hybridoma cell secreting monoclonal antibody against Yersinia pestis antigen, a hybridoma cell line and antibody which are obtained according to the method, and application thereof. The method comprises the following steps of: mouse immunization, cell fusion and inoculation, plague Fe antibody detection, and hybridoma cell clone. A hybridoma cell line secreting high titer monoclonal antibody against Yersinia pestis F1 antigen can continuously produce the monoclonal antibody against the Yersinia pestis F1 and provides an antibody source for establishing a quick plague diagnosis method.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a monoclonal antibody hybridoma cell line and a preparation method thereof. A hybridoma cell line 1B of the present invention 12 The strain was preserved at the China Type Culture Collection Center located in Wuhan, China on September 16, 2010, and its preservation number is CCTCC C201088; another hybridoma cell strain 2C of the present invention 6 The strain was preserved on September 16, 2010 at the China Center for Type Culture Collection located in Wuhan, China, and its preservation number is CCTCC C201089. Background technique [0002] Plague is a natural foci disease caused by Yersinia pestis. It is highly contagious and has a high fatality rate. In recent years, the world's plague epidemic has been on the rise, and the World Health Organization has listed plague as one of the re-epidemic infectious diseases. [0003] Due to the serious harm of plague, any misdiagnos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/12G01N33/577C12R1/91
Inventor 尹家祥董兴齐于国林杜春红唐雪钟佑宏陈平王鹏
Owner 云南省地方病防治所
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