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Bactrian camel heavy-chain (HC) variable-domain antibody resisting porcine circovirus 2 as well as preparation method and application thereof

A technology of porcine circovirus and single-domain heavy chain antibody, which is applied in the fields of virus/phage, antiviral immunoglobulin, botany equipment and methods, etc.

Active Publication Date: 2014-03-12
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Using Bactrian camels to prepare PCV2VHH immune antibody library, screen PCV2-specific VHH antibodies, and develop rapid pathogen diagnosis methods have not been reported at home and abroad.

Method used

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  • Bactrian camel heavy-chain (HC) variable-domain antibody resisting porcine circovirus 2 as well as preparation method and application thereof
  • Bactrian camel heavy-chain (HC) variable-domain antibody resisting porcine circovirus 2 as well as preparation method and application thereof
  • Bactrian camel heavy-chain (HC) variable-domain antibody resisting porcine circovirus 2 as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1 Preparation of Bactrian camel VHH heavy chain antibody against porcine circovirus type 2

[0057] 1. Materials and methods

[0058] 1.1 Materials

[0059] 1.1.1 Test animals, antigens, adjuvants

[0060] 1 male Bactrian camel (1 year old), 1 female Bactrian camel (6 months old); PCV2 inactivated vaccine (Ingelvac circoFLEX, Serial NO.309-211), recombinant PCV2 VLPs antigen (Shuanghui Yin, Shiqi Sun, Shunli Yang, et al. Self-assembly of virus-like particles of porcine circovirus type 2capsid protein expressed from Escherichia coli [J.] Virology Journal 2010,7:166).

[0061] 1.1.2 Strains, helper phages and reagents

[0062] Escherichia coli TG1 and HB2151, helper phage M13K07, and carrier pHEN2 were purchased from Novagen; expression vector pSMK was donated by Dr. Sun Shiqi from our institute; restriction enzymes were purchased from NEB; lymphocyte separation solution (1.077) was purchased from Shanghai Sangong; Trizol, DNA fragment recovery kit and plasmid...

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Abstract

The invention discloses a bactrian camel heavy-chain (HC) variable-domain antibody resisting a porcine circovirus 2 as well as a preparation method and an application thereof, and belongs to the field of a biotechnology. A PCV2 (Porcine Circovirus Virus) inactivated vaccine immunized bactrian camel is firstly utilized and a phage display technology is utilized to establish a PCV2 immune VHH antibody base; and a high-specificity variable domain antibody of heavy chains of HCAbs (VHH) which has a good affinity with the PCV, particularly a PCV2 virus antigen; and the antibody disclosed by the invention has an amino acid sequence shown as SEQ (sequence) ID NO.1 or an amino acid sequence which is shown as SEQ ID NO.2 and has the sequence isogeny being more than 95%. The PCV2 heavy-chain variable-domain antibody disclosed by the invention has the very important meanings on researching porcine circovirus, particularly researching a porcine circovirus 2-type infection mechanism and establishing rapid antigen detection with high sensitivity and specificity or diagnosing a reagent.

Description

technical field [0001] The present invention relates to an antibody and its preparation method and application, in particular to an anti-PCV2 Bactrian camel single domain heavy chain antibody. In addition, it also relates to the preparation method, function identification and application of the antibody. The invention belongs to the field of biotechnology. Background technique [0002] Porcine circovirus disease is a viral infectious disease of pigs that has become popular in recent years. The pathogen is porcine circovirus type 2 (PCV2). PCV2 is the main pathogen that causes multisystemic wasting syndrome in weaned piglets, and is also related to the occurrence of porcine reproductive disorders, porcine respiratory syndrome, porcine dermatitis and nephritic syndrome, porcine proliferative necrotizing pneumonia, congenital tremor and other diseases. [0003] Porcine circovirus belongs to the Circoviridae family and the genus Circovirus. The virus has no envelope and is a s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C12N15/13C12N15/63C12N7/01G01N33/569C12Q1/70C12Q1/68C12R1/93
Inventor 刘湘涛杨顺利尹双辉尚佑军孙世琪田宏靳野吴锦艳陈妍张克山
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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