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Monoclonal antibody and antibody combination against foot-and-mouth disease type O virus and its application in the detection of virus antigen and antibody

A monoclonal antibody, virus antigen technology, applied in antiviral immunoglobulin, application, immunoassay, etc., can solve the problem of large batch difference of polyclonal antibody

Active Publication Date: 2020-06-30
北京标驰泽惠生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is well known that polyclonal antibodies target multiple antigenic sites, and the batches of polyclonal antibodies vary greatly

Method used

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  • Monoclonal antibody and antibody combination against foot-and-mouth disease type O virus and its application in the detection of virus antigen and antibody
  • Monoclonal antibody and antibody combination against foot-and-mouth disease type O virus and its application in the detection of virus antigen and antibody
  • Monoclonal antibody and antibody combination against foot-and-mouth disease type O virus and its application in the detection of virus antigen and antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] 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

[0117] 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. Please also refer to the published patent document CN105348386A for the specific method.

[0118] The acquisition of hybridoma cell lines comprises the following steps:

[0119] 1. Animal immunity

[0120] 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 μg / mL. If necessary, the virus antigen can be concentrated in an ultrafiltration tube to increase the ...

Embodiment 2

[0155] Embodiment 2, obtain the hybridoma cell of the monoclonal antibody of the O type (O / Mya98 / BY / 2010) virus that continuously, stably secretes anti-foot-and-mouth disease

[0156] In this embodiment, the hybridoma that continuously and stably secretes the monoclonal antibody against the foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus obtained by immunizing mice with the foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus as the immunogen cell line.

[0157] The methods for obtaining the hybridoma cell line and detecting the potency are the same as those in Example 1. Five hybridoma cells that can stably secrete monoclonal antibodies against foot-and-mouth disease type O (O / Mya98 / BY / 2010) virus were obtained, and they were numbered 1F2, 2B4, 5C3, 4H9, and 7F5 according to the position of cloning production.

[0158] The titer determination results of the cell culture supernatant are shown in Table 4, and the titer ranged from 1:10 to 1:120, indicating that the targ...

Embodiment 3

[0168] Embodiment 3, prepare the monoclonal antibody of anti-foot-and-mouth disease O type (O / GX / 09-7) virus in a large amount

[0169] 1. Mass preparation and purification of monoclonal antibodies

[0170] 1. A large amount of monoclonal antibody was prepared by the method of inducing monoclonal antibody in animals: adult BalB / c mice were selected, and pristane (immunosuppressant, which can cause inflammatory reaction) 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.

[0171] 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 t...

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Abstract

The invention discloses a monoclonal antibody and antibody combination for resisting a foot-and-mouth disease type O virus and application thereof in detection of antigens and antibodies of the virus, and provides a non-specific monoclonal antibody 4H9anti and antibody combination for the foot-and-mouth disease type O virus. The antibody combination comprises specific monoclonal antibodies 6D6anti and 1F2anti, a non-specific monoclonal antibody 2H10anti and the non-specific monoclonal antibody 4H9anti for the foot-and-mouth disease type O virus. The monoclonal antibody and antibody combination can realize detection of antigens and antibodies of the foot-and-mouth disease type O virus, can quickly detect multiple type O virus strains, have favorable broad-spectrum activity in detection of the foot-and-mouth disease type O virus, have the characteristics of high specificity, high sensitivity and high detection speed, and can be used for production of foot-and-mouth disease type O virus vaccines, quality control of related products and evaluation of immune effects of the vaccines.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a monoclonal antibody of foot-and-mouth disease type O virus, a hybridoma cell line secreting the antibody, and the combination and application of the antibody and other antibodies. The application is embodied in the detection of the antigen and antibody of foot-and-mouth disease type O virus , to realize the monitoring of raw materials, intermediate products and finished products during the vaccine production process, as well as the monitoring of serum antibody levels after immunization with the foot-and-mouth disease O-type vaccine. Background technique [0002] Foot-and-mouth disease is an infectious disease common to cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV) infection. FMDV currently has seven serotypes: O, A, C, SAT1, SAT2, SAT3 (namely, South African foot-and-mouth disease virus types 1, 2, and 3) and Asia1 (Asian type 1). There is almost no immune prot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C12N15/13C12N5/20G01N33/577G01N33/569G01N33/68
CPCC07K16/1009C07K2317/56G01N33/56983G01N33/6854G01N2333/09G01N2469/20
Inventor 郑金来吴园园
Owner 北京标驰泽惠生物科技有限公司
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