Process for preparing chicken colibacillus pilus monoclonal antibody

A technology of monoclonal antibody and chicken Escherichia coli, which is applied in the direction of bacteria and fermentation, and can solve the problems of difficult standardization, large differences between batches, and low titer

Inactive Publication Date: 2004-03-31
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are many researches on F1 and P pili at home and abroad, but the research is greatly limited.
The main reason is that the pili antiserum used in current research is polyclonal antiserum, which has poor specificity, cross-reactivity, low titer, large batch-to-batch variation, troublesome preparation, difficult promotion, difficult standardization, and inability to effectively distingui

Method used

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  • Process for preparing chicken colibacillus pilus monoclonal antibody
  • Process for preparing chicken colibacillus pilus monoclonal antibody
  • Process for preparing chicken colibacillus pilus monoclonal antibody

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Embodiment Construction

[0089] Embodiments of the present invention are described below in conjunction with accompanying drawings:

[0090] figure 1The preparation process of the monoclonal antibody of the embodiment of the present invention is described. It can be seen from the figure that the splenic lymphocytes of BALB / C mice immunized with predetermined antigens are fused with myeloma cells (SP2 / 0) which can grow unrestricted in vitro to form hybridoma cells with parental characteristics. The hybridoma cells can not only synthesize and secrete antibodies against predetermined antigens like splenic lymphocytes, but also live forever in vitro like myeloma cells. A hybridoma cell line from a single cell can be obtained by cloning, and the antibody secreted by it is directed against the same antigenic determinant and is molecularly homogeneous, the so-called monoclonal antibody (Monoclonal antibody), referred to as monoclonal antibody .

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Abstract

A method of preparing monoclonal antibody against pillis of bacillus coli. It comprises: 1) screening good strains of bacillus coli; 2) preparing immunogens;3) immunizing BALB/C mice;4)fusing hybridoma cells; 5) screening hybridoma cells; 6) screening positive hybridoma cells;7) clonizing;8) clonizing again;9) inducing ascitic fluid containing monoclonal antibody; 10) measuring valence of monoclonal antibody in ascitic fluid. The monoclonal antibody has a high valence, could discriminate F1,P and other pillis, which could be used to determine pathogenic bacillus coli strains.

Description

technical field [0001] The invention belongs to the technical field of biological cell engineering, and in particular relates to a preparation method of chicken Escherichia coli pili monoclonal antibody. Background technique [0002] Chicken colibacillosis is a contact infectious disease caused by pathogenic Escherichia coli, and it is one of the most serious infectious diseases that endanger the chicken industry. There are more than 170 serotypes of chicken Escherichia coli, but most of them are non-pathogenic, and only a few are pathogenic bacteria. Its pili play an important role in the process of pathogenic Escherichia coli invading the body, colonizing the body, proliferating, and causing disease. There are many types of Escherichia coli pili and they are complex. But for chicken pathogenic Escherichia coli, only two important pili, F1 and P, have been found so far. Both pili are coded by the chromosome of the bacterium, and their molecular wei...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/08
Inventor 杜元钊董国雄范根成
Owner YEBIO BIOENG OF QINGDAO
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