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Anti-rabbit hemorrhagic disease virus VP60 albumen monoclonal antibody

A rabbit hemorrhagic virus, monoclonal antibody technology, applied in antiviral immunoglobulin, biochemical equipment and methods, microorganisms, etc., can solve the problems of unfavorable RHDV monoclonal antibody application, low monoclonal antibody titer, etc. The preparation method is simple and feasible, the preparation method is simple, and the specificity is strong

Inactive Publication Date: 2009-09-02
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The titer of monoclonal antibody prepared by using purified RHDV virus as immunogen is usually low, which is not conducive to the application of RHDV monoclonal antibody

Method used

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  • Anti-rabbit hemorrhagic disease virus VP60 albumen monoclonal antibody
  • Anti-rabbit hemorrhagic disease virus VP60 albumen monoclonal antibody
  • Anti-rabbit hemorrhagic disease virus VP60 albumen monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0026] (1) Construction of recombinant VP60 protein expression vector

[0027]Referring to the genome sequence of RHDV of Jilin strain in China (AY523410) in GenBank, a pair of primers were designed by Primer 5.0 software. Upstream primer P1: 5'-ATA GTCGAC ATG GAG GGC AAA G-3'; Downstream primer P2: 5'-GCC TCG AGT CAG ACA TAA GM AAG CCA-3'. A Sal I restriction site was added to the 5' end of the upstream primer, and a Xho I restriction site was introduced to the 5' end of the downstream primer. Amplify RHDV capsid protein VP60 gene with RT-PCR method (Wang Fang etc., expression of capsid protein of rabbit haemorrhagic disease virus in insect cells and immune protection effect to rabbits, Journal of Animal Husbandry and Veterinary Medicine, 2008, 39 (10) , 1382~1387), connected to the donor plasmid pFastBac after enzyme digestion TM 1 (purchased from Invitrogen), transformed DH5α competent cells, extracted the plasmid and cut it with Sal I and Xho I, and obtained the recombin...

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Abstract

The invention relates to an anti-rabbit hemorrhagic disease virus (RHDV)VP60 albumen monoclonal antibody, and belongs to the technical field of biology. An SP2 / 0 myeloma cell and a BALB / c mouse splenic cell immunized by utilizing RHDV to recombine VP60 albumen undergo cell fusion, are selectively cultured by an HAT culture medium and undergo double ELISA screening by utilizing the recombined VP60 albumen and RHDV; the obtained cell culture supernatant is checked up and screened respectively to obtain a hybrid tumor cell strain A3C which can stably excrete the anti-RHDV VP60 albumen monoclonal antibody; the ascitic fluid ELISA titer of the A3C is detected to be 1:327,600; and according to identification, the monoclonal antibody can specifically combine the expressed recombined VP60 albumen as well as the RHDV, and one single reaction strip appears in both specific combinations, thereby proving that the anti-RHDV VP60 albumen monoclonal antibody is a VP60 specific antibody of the RHDV capsid albumen.

Description

technical field [0001] The invention relates to an anti-rabbit hemorrhagic disease virus (RHDV) VP60 protein monoclonal antibody, which belongs to the field of biotechnology and relates to antibody engineering technology, in particular to an anti-rabbit hemorrhagic disease virus (RHDV) VP60 protein monoclonal antibody. Background technique [0002] Rabbit hemorrhagic disease (RHD), also known as rabbit plague, is a rabbit infectious disease characterized by acute, highly contagious, and large-scale death caused by rabbit hemorrhagic disease virus (RHDV). Infected rabbits usually die within 48 to 72 hours. The main features are highly infectious, respiratory system hemorrhage, liver necrosis, visceral edema, congestion and hemorrhage and other changes, which have caused huge economic losses to the rabbit industry all over the world. The International Veterinary Bureau listed the disease as a Class B infectious disease, and the No. 96 Order of the Ministry of Agriculture of my...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12P21/08C12N5/20
Inventor 王芳范志宇胡波蔡少平徐为中张则斌何孔旺
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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