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A kind of whole pig origin African swine fever virus monoclonal antibody and preparation method thereof

An African swine fever virus and monoclonal antibody technology, which is applied in the biological field to achieve the effects of strong specificity, good antibody affinity and reduced production costs

Active Publication Date: 2021-12-14
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0004] The invention patents with publication numbers CN112592401A and CN112500478A respectively disclose a ScFv antibody VH-VLλ11 and VH-VLλ6 against African swine fever virus, and their preparation methods, although VH-VLλ11 and VH-VLλ6 are antibodies to African swine fever virus , but the antibodies are all single-chain antibodies, single-chain antibodies only include the heavy chain variable region and the light chain variable region, not including the constant region

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  • A kind of whole pig origin African swine fever virus monoclonal antibody and preparation method thereof
  • A kind of whole pig origin African swine fever virus monoclonal antibody and preparation method thereof
  • A kind of whole pig origin African swine fever virus monoclonal antibody and preparation method thereof

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Embodiment Construction

[0041] In the following, the present invention will be described in more detail through specific embodiments, so as to facilitate the understanding of the technical solution of the present invention, but it is not used to limit the protection scope of the present invention.

[0042] 1. Sample collection and PBMC isolation

[0043] 10ml of anticoagulated blood from an African swine fever immunized pig (5 months old, from a pig farm in Xinxiang, Henan) was collected from the jugular vein for PBMC isolation. The operation method adopts the method described in the instruction manual of the porcine peripheral blood lymphocyte isolation kit (Tianjin Haoyang / TBD, LTS1110).

[0044] 2. Isolation and screening of target B lymphocytes

[0045] ASFV antigen-specific porcine B lymphocytes were selected by dual labeling of ASFV and anti-pig IgG. After inactivation of ASFV, biotin (Thermo Scientific TM EZ-Link TM Sulfo-NHS-LC-Biotinylation Kit, Thermofish, 21435) for labeling.

[0046]...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a monoclonal antibody of whole swine origin African swine fever virus and its preparation method and application. The monoclonal antibody comprises a heavy chain constant region, a heavy chain variable region, a light chain constant region and a light chain variable region, and the amino acid sequence of the heavy chain variable region is as SEQ ID NO: 1 or SEQ ID NO: 5 As shown, the amino acid sequence of the light chain variable region is shown in SEQ ID NO:2 or SEQ ID NO:6. The whole pig-derived monoclonal antibody specific for African swine fever virus of the present invention retains the natural pairing of light chain and heavy chain variable regions, and has good gene diversity, whole pig origin, good antibody affinity, strong specificity, etc. Advantages, high-efficiency expression can be performed in eukaryotic expression system. The invention provides new raw materials for the development of African swine fever virus diagnostic kits, and also provides new technical support for the development of preventive and therapeutic drugs for whole pig-derived monoclonal antibodies.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a monoclonal antibody of whole swine origin African swine fever virus and a method for preparing the monoclonal antibody by high-throughput single B cell PCR technology. Background technique [0002] African swine fever (ASF) is an acute and highly contagious disease of pigs caused by African swine fever virus (ASFV), with a short course and a fatality rate of up to 100%. The World Health Organization (OIE) lists it as a legally notifiable animal disease, and my country lists it as a first-class animal disease. ASF was first discovered in Kenya in 1921. It was introduced into my country in August 2018, and then quickly spread to many provinces and cities in my country, causing huge economic losses to the development of my country's pig industry. So far, there is no effective commercial vaccine for the prevention and control of ASF. In the short term, early diagnosis and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C12N15/13C12N15/85G01N33/577G01N33/569A61K39/42A61P31/20
CPCC07K16/081C12N15/85G01N33/577G01N33/56983A61P31/20C07K2317/52C07K2317/56G01N2333/01
Inventor 郑海学曹丽艳田宏王琦孔祥雨宋锐石正旺
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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