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Bactrian camel VHH (variable domain of the heavy chain of HACbs) heavy-chain antibody for resisting foot-and-mouth disease AsiaI type viruses, preparation method and use thereof

A technology of heavy chain antibody and Bactrian camel, applied in antiviral immunoglobulin, biochemical equipment and methods, recombinant DNA technology, etc.

Active Publication Date: 2012-10-03
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using Bactrian camels to prepare the Asia I type immune antibody library, so as to screen and obtain the VHH heavy chain antibody that can specifically bind to the foot-and-mouth disease Asia I type virus antigen, to develop a rapid diagnosis method for pathogens has not been reported at home and abroad.

Method used

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  • Bactrian camel VHH (variable domain of the heavy chain of HACbs) heavy-chain antibody for resisting foot-and-mouth disease AsiaI type viruses, preparation method and use thereof
  • Bactrian camel VHH (variable domain of the heavy chain of HACbs) heavy-chain antibody for resisting foot-and-mouth disease AsiaI type viruses, preparation method and use thereof
  • Bactrian camel VHH (variable domain of the heavy chain of HACbs) heavy-chain antibody for resisting foot-and-mouth disease AsiaI type viruses, preparation method and use thereof

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Experimental program
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Effect test

Embodiment 1

[0039] 1. Materials and methods

[0040] 1.1 Materials

[0041] 1.1.1 Test animals, antigens, adjuvants and AsiaI antibody detection kits

[0042]1 male Bactrian camel (1 year old), 1 female Bactrian camel (6 months old); recombinant AsiaI / YNBS / 58 strain VLPs antigen, AsiaI / YNBS / 58 (full genome sequence of foot-and-mouth disease virus AsiaI / YNBS / 58 strain Analysis, Chang Huiyun et al., Acta Virus, 2007(5):407-411) Whole virus inactivated antigen; 206 adjuvant; FMDV Asia Type I antibody liquid phase ELISA detection kit was developed and provided by Lanzhou Veterinary Research Institute.

[0043] 1.1.2 Strains, helper phages and reagents

[0044] Escherichia coli TG1 and HB2151, helper phage M13K07, and vector pHEN2 were donated by Dr. Wang Xiangbin.

[0045] Expression vector pE-Sumo (Kanr, LifeSensors, Inc, vector map as Figure 7 shown. ), restriction endonuclease was purchased from NEB, lymphocyte separation fluid (1.077) was purchased from Shanghai Sangong, Trizol, nic...

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Abstract

The invention discloses a bactrian camel VHH (variable domain of the heavy chain of HACbs) heavy-chain antibody for resisting foot-and-mouth disease AsiaI type viruses, a preparation method and use thereof, belonging to the field of biotechnology. In the method, recombinant AsiaI type virus-like particle antigens are used for immunizing bactrian camels; an AsiaI VHH immune antibody library is established with phage display technology; high-specificity single-domain antibodies having good affinity with foot-and-mouth disease AsiaI type virus antigens are screened; the antibody provided by the invention has amino acid sequence shown by SEQ ID NO.1 or amino acid sequence which has sequence homology more than 98% with the sequence shown by SEQ ID NO.1. The AsiaI VHH heavy-chain antibody provided by the invention has great importance for researching AsiaI type virus infection mechanisms and producing high-sensitive and specific antigen fast detection or diagnosis reagents.

Description

technical field [0001] The invention relates to an antibody and its preparation method and application, in particular to a Bactrian camel VHH heavy chain antibody against foot-and-mouth disease AsiaI virus. In addition, the present invention also relates to the preparation method, functional identification and application of the antibody. The invention belongs to the field of biotechnology. Background technique [0002] Foot-and-Mouth Disease (FMD) is caused by Foot-and-Mouth Disease Virus (FMDV) and is an acute, febrile, highly contactable, virulent animal infectious disease mainly caused by cloven-hoofed animals. , is an animal infectious disease that threatens the development of animal husbandry in the world. Foot-and-mouth disease is highly contagious and highly morbid, and the lethality rate to young animals can sometimes reach 100%, so it is listed as the first among the 14 A-class infectious diseases of animals by the World Organization for Animal Health (OIE). my c...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/11C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12Q1/70C12Q1/68G01N33/569
Inventor 刘湘涛尹双辉杨顺利尚佑军孙世琪靳野田宏周广清
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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