中国动物传染病学报2024，Vol.32Issue(6)：176-181,6.
非洲猪瘟病毒p34蛋白单克隆抗体的制备及鉴定
Preparation and Characterization of a Monoclonal Antibody Against p34 Protein of African Swine Fever Virus
摘要
Abstract
African swine fever(ASF)is a highly contagious disease caused by African swine fever virus(ASFV)infection in domestic pigs and wild boars.ASFV encodes more than 150 proteins,of which most proteins remain to be characterized.This study aimed to express ASFV p34 protein and prepare monoclonal antibodies(mAbs)for the functional study.Firstly,the recombinant prokaryotic expression plasmid PET-21a-p34 was constructed and transformed into competent E.coli BL21(DE3)cells.The soluble His-p34 recombinant protein was expressed with IPTG induction and identified by SDS-PAGE and Western blot.The purified protein was used to immunize the mice.As a result,a hybridoma cell line capable of stably secreting the mAbs against p34 protein was obtained by cell fusion and ELISA screening.Western blot analysis showed that the mAb specifically recognized exogenous p34 protein overexpressed by transfected plasmid and endogenous p34 protein expressed by ASFV-infected porcine alveolar macrophages(PAMs).Immunofluorescence assay showed that both exogenous transfected and endogenous expressed p34 proteins were specifically labeled for visual reaction.Immunoprecipitation(IP)assay showed that the mAb specifically bound to exogenous p34 protein in HEK293T cells and endogenous p34 protein in ASFV-infected PAMs.In conclusion,the mAb to p34 protein was successfully prepared in this study,which could be used for the identification and functional study of endogenous p34 protein.关键词
非洲猪瘟病毒/p34蛋白/原核表达/单克隆抗体Key words
African swine fever virus/p34 protein/prokaryotic expression/monoclonal antibody引用本文复制引用
王晓,刘传霞,鲍苗菲,李雪雯,李婷婷,陈欣,郑君,翁长江..非洲猪瘟病毒p34蛋白单克隆抗体的制备及鉴定[J].中国动物传染病学报,2024,32(6):176-181,6.基金项目
十四五国家重点研发计划项目(2021YFD1800100) （2021YFD1800100）
国家自然科学基金项目(32172874) （32172874）
