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A kind of scfv antibody against African swine fever virus and preparation method thereof

An African swine fever virus, African swine fever technology, applied in antiviral agents, antiviral immunoglobulins, botanical equipment and methods, etc., to achieve the effect of strong specificity and small immunogenicity

Active Publication Date: 2020-11-03
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

[0004] Looking back at the development of the antibody field in recent years, we can see that traditional antibodies can no longer meet the needs of the current antibody market. People are in urgent need of a flexible antibody that is easy to prepare, strong in specificity, high in stability, and small in molecular weight. Efficiently act on specific parts

Method used

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  • A kind of scfv antibody against African swine fever virus and preparation method thereof
  • A kind of scfv antibody against African swine fever virus and preparation method thereof
  • A kind of scfv antibody against African swine fever virus and preparation method thereof

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Experimental program
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Effect test

Embodiment

[0068] Example Preparation of Anti-African Swine Fever Virus ScFv Antibody

[0069] 1. Materials and methods

[0070] 1.1 Materials

[0071] 1.1.1 Test animals

[0072] An African swine fever immune resistant pig (5 months old, from a pig farm in Xinxiang, Henan).

[0073] 1.1.2 Strains and reagents

[0074] LTS1110 Porcine Peripheral Blood Lymphocyte Separation Kit was purchased from Tianjin Haoyang Biological Products Company; PMD TM 20-T carrier, E.coli JM109 competent, HS DNA Polymerase, RNase-free Water, DL2,000 DNA Marker, and 6×Loading Buffer were all purchased from Bao Biological Company; BL21(DE3) competent was purchased from Beijing Quanshijin; Trizol, nickel affinity chromatography resin, and DNA fragment recovery Kits and plasmid extraction kits are OMEGA products; molecular biology reagents are from Sigma; other biochemical reagents are of domestic analytical grade.

[0075] 1.2 Method

[0076] 1.2.1 Lymphocyte separation

[0077] Collect 100mL of pig ant...

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Abstract

The invention discloses an ScFv antibody for resisting African swine fever virus and a preparation method thereof, lymphocytes are separated from peripheral blood of a naturally infected African swinefever immune resistant pig, total mRNA of the separated lymphocytes is extracted, a total cDNA fragment is obtained through an RT-PCR method, by taking the cDNA as a template, under the action of a corresponding primer with a Linker linker, a swine-derived ScFv antibody genetic sequence is obtained through an SOE-PCR method, the swine-derived ScFv antibody genetic sequence is connected with a PET-30a vector, and then transforms BL21 competent cells, ScFv antibody aiming at the African swine fever virus is screened from a transformed single colony, and the screened ScFv antibody is subjected to preliminary activity identification by adopting an ELISA (Enzyme-Linked Immuno Sorbent Assay) test, so that the antibody has the African swine fever reaction activity. The invention provides a new material for early diagnosis and prevention and control of African swine fever, and provides a new technical support for controlling epidemic transmission of African swine fever as soon as possible.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an anti-African swine fever virus ScFv antibody and a preparation method thereof. Background technique [0002] African swine fever (African swine fever, ASF) is a highly fatal hemorrhagic disease of pigs caused by African swine fever virus (African swine fever Virus, ASFV), which is highly contagious. ASFV is an icosahedral double-stranded DNA virus with a diameter of 200 nm. It is the only member of the African swine fever virus family and the only known vector-borne virus. African swine fever first broke out in Kenya, Africa in 1921, and was introduced to my country in August 2018. As of February 2019, African swine fever has occurred in 24 provinces across the country, causing huge economic losses to the pig industry. At present, there is no effective vaccine against this virus in the world, so the early diagnosis of African swine fever plays a vital role in the prev...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C12N15/13C12N15/70A61K39/42A61P31/20G01N33/569
CPCA61K2039/505A61P31/20C07K16/081C07K2317/622C12N15/70G01N33/56983
Inventor 郑海学王丽娟田宏石正旺罗俊聪杨波张克山茹毅李丹杨帆
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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