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Hybridoma cell strain, foot-and-mouth disease resistant O-type (O/GX/09-7) virus specificity monoclonal antibody secreted by hybridoma cell strain and application of hybridoma cell strain and antibody to detection of foot-and-mouth disease O-type viruses

A hybridoma cell line, monoclonal antibody technology, applied in the biological field, can solve problems such as insufficient application detection

Active Publication Date: 2016-02-03
JINYUBAOLING BIO PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are occasional studies on the preparation of FMD monoclonal antibodies in China, but they are not enough to be applied to the detection of FMD O (O / GX / 09-7) virus, which can specifically detect FMD O (O / GX / 09-7 ) virus monoclonal antibody is still a research direction

Method used

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  • Hybridoma cell strain, foot-and-mouth disease resistant O-type (O/GX/09-7) virus specificity monoclonal antibody secreted by hybridoma cell strain and application of hybridoma cell strain and antibody to detection of foot-and-mouth disease O-type viruses
  • Hybridoma cell strain, foot-and-mouth disease resistant O-type (O/GX/09-7) virus specificity monoclonal antibody secreted by hybridoma cell strain and application of hybridoma cell strain and antibody to detection of foot-and-mouth disease O-type viruses
  • Hybridoma cell strain, foot-and-mouth disease resistant O-type (O/GX/09-7) virus specificity monoclonal antibody secreted by hybridoma cell strain and application of hybridoma cell strain and antibody to detection of foot-and-mouth disease O-type viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1, obtain the hybridoma cell of the monoclonal antibody of continuous, stably secreting anti-foot-and-mouth disease O type (O / GX / 09-7) virus

[0071] In this embodiment, the hybridoma that continuously and stably secretes monoclonal antibodies against foot-and-mouth disease type O (O / GX / 09-7) virus is obtained by immunizing mice with foot-and-mouth disease type O (O / GX / 09-7) virus cell line.

[0072] The acquisition of hybridoma cell lines comprises the following steps:

[0073] 1. Animal immunity

[0074] 1) Basic immunity: using foot-and-mouth disease (O / GX / 09-7) virus (provided by Jinyu Baoling Biopharmaceutical Co., Ltd.) as the immunogen (antigen), it was determined by sucrose density gradient centrifugation, with a purity of ≥85% and a concentration of 10 -(1000 (g / mL,. If necessary, the virus antigen can be concentrated in an ultrafiltration tube to increase the virus content. After concentration, the antigen is mixed with Freund's complete adjuvant ...

Embodiment 2

[0112] Embodiment 2, prepare the monoclonal antibody of anti-foot-and-mouth disease O type (O / GX / 09-7) virus in a large amount

[0113] 1. Mass preparation and purification of monoclonal antibodies

[0114] 1. A large amount of monoclonal antibodies were prepared by inducing monoclonal antibodies in animals: adult BALB / c mice were selected, and pristine (immunosuppressant, which can cause inflammatory reactions) was inoculated intraperitoneally, 0.5 mL per mouse. After 7-10 days, the 16th generation hybridoma cells 6D6 or 2H10 were inoculated intraperitoneally, 2×10 per mouse 6 -3×10 6 indivual. After an interval of 5 days, when the abdomen is obviously enlarged and the skin feels tense when touched with hands, the ascites can be collected with a No. 9 needle.

[0115] 2. Antibody purification: Centrifuge the ascitic fluid (10000r / min for 30 minutes), remove cell components and other precipitates, and collect the supernatant. The supernatant was washed with 15-30 times the...

Embodiment 3

[0128] Embodiment 3, detect foot-and-mouth disease type O (O / GX / 09-7) virus with monoclonal antibody 6D6anti, 2H10anti and ELISA double antibody sandwich method

[0129] 1. Detection of different concentrations of foot-and-mouth disease type O (O / GX / 09-7) virus with monoclonal antibody 6D6anti and ELISA double-antibody sandwich method

[0130] 1. 6D6anti is both a coated antibody and a labeled antibody to detect different concentrations of foot-and-mouth disease type O (O / GX / 09-7) virus——6D6anti (coated)-6D6anti (labeled) pairing kit

[0131] Documents (Number and molecular weights of Food-and-Mouth Disease Virus Capsid Proteins and the Effects of Maleylation, Journal of Virology, 1971, Vol7, No.2, P250-259) record that the membrane proteins (VP1, VP2, VP3 and VP4) of foot-and-mouth disease virus have multiple copies, so the monoclonal antibody 6D6anti Both the coated antibody and the labeled antibody are used to specifically detect the foot-and-mouth disease type O (O / GX / 09-7...

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Abstract

The invention discloses a hybridoma cell strain 6D6 of a specificity monoclonal antibody capable of continuously and stably secreting foot-and-mouth disease resistant O-type (O / GX / 09-7) viruses, and the specificity monoclonal antibody secreted by the hybridoma cell strain. The 6D6anti can recognize the foot-and-mouth disease O-type (O / GX / 09-7) viruses in a specificity mode, the 6D6anti is used as a coating and an enzyme labeled antibody in ELISA detection and can be used for detecting the foot-and-mouth disease O-type (O / GX / 09-7) viruses, and the advantages of high specificity and high sensitivity are achieved; the hybridoma cell strain, the antibody and the application can play an important role in detection of the foot-and-mouth disease O-type (O / GX / 09-7) viruses, vaccine production and epidemiologic study.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a hybridoma cell strain and a monoclonal antibody secreted therefrom, in particular to a specific monoclonal antibody to foot-and-mouth disease type O (O / GX / 09-7) virus and a hybridoma cell strain secreting the antibody And the application of the antibody in detecting foot-and-mouth disease type O (O / GX / 09-7) virus. Background technique [0002] Foot-and-mouth disease (FMD) belongs to a class of infectious diseases. It is a sexual and contact infectious disease characterized by blisters on the oral mucosa, hooves and breast skin. Foot-and-mouth disease virus mainly infects artiodactyls, and is occasionally seen in humans and other animals. Its main hosts include pigs, cattle, sheep and other major domestic animals and other domestic and wild artiodactyls. There are more than 70 susceptible animals, and the most susceptible animals are Cattle, pigs, camels, sheep, deer, wild boars, bi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/577G01N33/569G01N33/558G01N33/543G01N33/535C12R1/91
Inventor 刘国英郑金来范秀丽吴园园任旭荣李蓉张燕红郝金宝魏学峰商晓桂
Owner JINYUBAOLING BIO PHARMA CO LTD
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