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A kind of anti-African swine fever virus scfv antibody vh-vlλ11 and preparation method thereof

An African swine fever virus, VH-VL technology, applied in the direction of antiviral agents, antiviral immunoglobulins, microorganism-based methods, etc., can solve problems that have not been disclosed yet

Active Publication Date: 2021-07-20
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

ScFv antibodies against African swine fever virus whose light chain is other than κ light chain have not been disclosed in the prior art

Method used

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  • A kind of anti-African swine fever virus scfv antibody vh-vlλ11 and preparation method thereof
  • A kind of anti-African swine fever virus scfv antibody vh-vlλ11 and preparation method thereof
  • A kind of anti-African swine fever virus scfv antibody vh-vlλ11 and preparation method thereof

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Embodiment Construction

[0097] The present invention is described in detail below through specific examples, but the scope of protection of the present invention is not limited to the following examples, anyone skilled in the art on the basis of the present invention, in combination with the technical solutions conceivable by common knowledge in the field, All belong to the protection scope of the present invention.

[0098] 1. Preparation of Anti-African Swine Fever Virus ScFv Antibody VH-VLλ11

[0099] 1. Materials and methods

[0100] 1.1 Materials

[0101] 1.1.1 Test animals

[0102] An African swine fever immunized pig (5 months old, from a pig farm in Xinxiang, Henan).

[0103] 1.1.2 Strains and reagents

[0104] 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 Biolo...

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Abstract

The invention relates to a ScFv antibody VH‑VLλ11 against African swine fever virus and a preparation method thereof. The amino acid sequence of the heavy chain variable region of the ScFv antibody is shown in SEQ ID NO:1, and the amino acid sequence of the light chain variable region of the ScFv antibody is shown in SEQ ID NO:2. The present invention isolates lymphocytes from the peripheral blood of naturally infected African swine fever immunized and resistant pigs, extracts the total mRNA of the isolated lymphocytes, and obtains the total cDNA fragments by RT-PCR, using the cDNA as a template, in the corresponding Under the action of the primers of the Linker adapter, the porcine ScFv antibody gene sequence was obtained by SOE‑PCR, and the ScFv antibody gene sequence was constructed into a PET‑30a vector, transformed into BL21 (DE3) competent cells, and screened to obtain a gene sequence targeting African pigs. The ScFv antibody VH‑VLλ11 of pestivirus, the preliminary activity identification of the ScFv antibody VH‑VLλ11 obtained by screening found that the antibody has African swine fever reactivity. The proposal of the present invention provides new materials for the early diagnosis and prevention and control of African swine fever, and provides new technical support for controlling the epidemic spread 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 VH-VLλ11 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...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C12N15/13C12N15/70C12N1/21G01N33/569A61K39/42A61P31/20C12R1/19
CPCA61K2039/505A61P31/20C07K16/081C07K2317/56C07K2317/622C07K2317/76C12N15/70G01N33/56983G01N2333/01
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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