Anti-African swine fever virus ScFv antibody VH-VL lambda 6 and preparation method thereof

Abstract

The invention relates to an anti-African swine fever virus ScFv antibody VH-VL lambda 6 and a preparation method thereof. The amino acid sequence of a heavy chain variable region of the ScFv antibodyVH-VL lambda 6 is shown as SEQ ID NO: 1, and the amino acid sequence of a light chain variable region of an ScFv antibody is shown as SEQ ID NO: 2. Lymphocyte is separated from peripheral blood of a naturally infected African swine fever immune tolerant pig, total mRNA of the lymphocyte is extracted and separated, a total cDNA fragment is obtained by an RT-PCR method, cDNA serves as a template, aswine-derived ScFv antibody gene sequence is obtained by an SOE-PCR method under the action of a corresponding primer with a Linker connector, and the ScFv antibody gene sequence is constructed to a PET-30a vector. BL21 (DE3) competent cells are converted, screening is performed to obtain the ScFv antibody VH-VL lambda 6 aiming at the African swine fever virus, and preliminary activity identification is performed on the screened ScFv antibody VH-VL lambda 6 to find that the antibody has African swine fever reaction activity. The invention provides a new material for early diagnosis, preventionand control of the African swine fever, and provides a new technical support for controlling epidemic situation propagation of the 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-VLλ6 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 ...

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

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