抗非洲猪瘟病毒单链抗体的构建、表达及活性鉴定

doi:10.11843/j.issn.0366-6964.2021.05.018

Abstract

Abstract: In this study, we aimed to prepare anti-ASFV pig-derived single-chain antibody (ScFv), and identify its biological activity, screen out the pig-derived ScFv with ASFV reactivity, and provide a new diagnosis for ASFV material. the peripheral blood of ASFV-infected and recovered pigs were collected to separate lymphocytes, then lymphocyte total RNAs were extracted and used as a template to obtain pig-derived IgG heavy chain variable region and light chain variable region genes by PCR amplification, using SOE-PCR technology to expand. The pig-derived ScFv gene was obtained by splicing; the pET-30a-ScFv expression vector was constructed, and the protein was expressed and purified. The reactivity of the ScFv antibody was identified by ELISA and IFA. The results showed that the pig-derived ScFv (VH-VLκ, VH-VLλ) genes were successfully amplified, and an scFv (VH-VLλ₁₁) antibody that was reactive with ASFV was identified. The results showed that an ScFv (VH-VLλ₁₁) antibody that was reactive with ASFV was successfully screened, providing new raw material for ASFV diagnosis, prevention and control.

Key words: ASFV, SOE-PCR, ScFv, diagnosis and prevention

CLC Number:

S852.659.1

WANG Lijuan, SHI Zhengwang, YANG Bo, MA Yuan, LUO Juncong, WAN Ying, SONG Rui, TIAN Hong, ZHENG Haixue. Construction, Expression and Activity Identification of Anti-ASFV Single-Chain Antibody[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(5): 1328-1336.

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Construction, Expression and Activity Identification of Anti-ASFV Single-Chain Antibody

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