Hybridoma cell strain and monoclonal antibody secreted by hybridoma cell strain for resisting foot-and-mouth disease O-type viruses and application

A technology of hybridoma cell lines and monoclonal antibodies, which is applied to hybridoma cell lines and the monoclonal antibodies secreted by them to detect foot-and-mouth disease type O virus. In the field of hybridoma cell lines, it can solve the problem of specific detection that has not yet been seen. monoclonal antibodies etc.

Active Publication Date: 2016-02-03
北京标驰泽惠生物科技有限公司 +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 no

Method used

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  • Hybridoma cell strain and monoclonal antibody secreted by hybridoma cell strain for resisting foot-and-mouth disease O-type viruses and application
  • Hybridoma cell strain and monoclonal antibody secreted by hybridoma cell strain for resisting foot-and-mouth disease O-type viruses and application
  • Hybridoma cell strain and monoclonal antibody secreted by hybridoma cell strain for resisting foot-and-mouth disease O-type viruses and application

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Experimental program
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Embodiment 1

[0069] Embodiment 1, obtain the hybridoma cell of the monoclonal antibody that continuously, stably secretes anti-foot-and-mouth disease O type (O / Mya98 / BY / 2010) virus

[0070] The method for obtaining hybridoma cell line 1F2 comprises the following steps:

[0071] 1. Animal immunity

[0072]1) Basic immunity: using foot-and-mouth disease (O / Mya98 / BY / 2010) virus (provided by Jinyu Baoling Bio-Pharmaceutical Co., Ltd.) as the immunogen, determined by sucrose density gradient centrifugation method, the purity is ≥85%, and the concentration is 10-1000 μg / 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 (purchased from Sigma Company) in equal volumes and fully emulsified, and the emulsion is injected subcutaneously at multiple points. Each Balb / c small Rats (8-12 weeks old, female, SPF-grade animal culture, purchased from the Experimental An...

Embodiment 2

[0107] Embodiment 2, prepare the monoclonal antibody of anti-foot-and-mouth disease O type (O / Mya98 / BY / 2010) virus in a large amount

[0108] 1. Mass preparation and purification of monoclonal antibodies

[0109] 1. Use the method of inducing monoclonal antibodies in animals to prepare a large amount of monoclonal antibodies: select adult BALB / c mice, and inoculate pristane intraperitoneally, 0.5 mL per mouse. After 7-10 days, the 16th generation hybridoma cell 1F2 was 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.

[0110] 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 volume of pH7.0-7.610mMPBS (recipe: NaH 2 PO 4 2H 2 O0.296g, Na ...

Embodiment 3

[0123] Embodiment 3, detect foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus with monoclonal antibody 1F2anti and ELISA double antibody sandwich method

[0124] 1. Detection of different concentrations of foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus with monoclonal antibody 1F2anti and ELISA double-antibody sandwich method

[0125] There are 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) to record that the membrane proteins (VP1, VP2, VP3 and VP4) of foot-and-mouth disease virus have multiple copies, so the present invention uses monoclonal The antibody 1F2anti is both a coating antibody and a labeled antibody to specifically detect the foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus.

[0126] Detect foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus with monoclonal antibody 1F2anti and ELISA double antibody sandwich method, detection...

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Abstract

The invention relates to a hybridoma cell strain and a monoclonal antibody secreted by the hybridoma cell strain for resisting foot-and-mouth disease O-type viruses and an application. The foot-and-mouth O-type (O/Mya98/BY/2010) virus capable of inducing the immune reaction of the body is used as immunogen to obtain the hybridoma cell strain 1F2 capable of sustainably and stably secreting the monoclonal antibodies for resisting the foot-and-mouth disease O-type (O/Mya98/BY/2010) virus, and the cell strain secretes the monoclonal antibody 1F2anti. The monoclonal antibody can specifically identify the foot-and-mouth disease O-type (O/Mya98/BY/2010) virus, can be used for detecting the foot-and-mouth disease O-type (O/Mya98/BY/2010) virus, has advantages of high specificity and high sensitivity and can be applied to the detection of the foot-and-mouth disease O-type (O/Mya98/BY/2010) virus, vaccine production and epidemiology research.

Description

technical field [0001] The invention belongs to the hybridoma cell line in the field of biotechnology and its secreted monoclonal antibody and application, in particular to the monoclonal antibody of foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus and the hybridoma cell secreting the antibody strain and the application of the antibody in the detection of foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus. Background technique [0002] Foot-and-mouth disease is an acute, febrile, and contagious infectious disease common to cloven-hoofed animals caused by Foot-and-mouth disease virus (FMDV) infection. The most susceptible animals are cattle, pigs, camels, sheep, deer, etc., wild boars Wild animals such as wild cattle and bison are also susceptible to the disease. Foot-and-mouth disease is endemic in Asia, Africa, the Middle East, and South America, and there are sporadic cases in non-endemic areas. [0003] After the onset of foot-and-mouth disease, it is generally ...

Claims

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Application Information

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IPC IPC(8): C12N5/20C12N15/13C07K16/10G01N33/577G01N33/569G01N33/558G01N33/543G01N33/535C12R1/91
Inventor 郑金来张翀宇吴园园刘国英任旭荣李蓉范秀丽郝金宝苍枫刘飞
Owner 北京标驰泽惠生物科技有限公司
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