河南农业科学2025，Vol.54Issue(2)：124-130,7.DOI:10.15933/j.cnki.1004-3268.2025.02.014
非洲猪瘟病毒D205R蛋白单克隆抗体的制备和抗原表位鉴定
Preparation and Epitope Analysis of Monoclonal Antibody against African Swine Fever Virus D205R Protein
摘要
Abstract
African swine fever(ASF)is a lethal infectious disease,and few vaccines or drugs were available to prevent or control ASF to date.In order to provide important materials for the diagnosis of African swine fever virus(ASFV),the ASFV recombinant D205R protein was expressed by E.coli system,a monoclonal antibody(mAb)against D205R protein was prepared,and antigenic epitope of mAb was identified.The results showed that the pET32a-D205R expression plasmid was successfully constructed,and the recombinant D205R protein with a size of about 44 ku was purified.Western blot assay showed that the recombinant D205R protein reacted specifically with ASFV-positive serum and had good immunogenicity.By hybridoma cell fusion and screening,mAb 19A5 was obtained.Western blot and indirect immunofluorescence assays showed that mAb 19A5 specifically recognized eukaryotically expressed recombinant D205R protein,and detected wild-type D205R protein.Alanine scanning identified 167-SDPPVVWLGGRPGD-180 as the antigenic epitope of mAb 19A5,and S167,W173,L174,G175,P178,and D180 were the key amino acids that binded to mAb 19A5.Homologous and structural analysis revealed that the antigenic epitope was highly conserved and located on the surface of the protein,which was a linear epitope.In conclusion,mAb 19A5 was successfully prepared and the antigenic epitope recognized by mAb 19A5 was identified.关键词
非洲猪瘟病毒/D205R蛋白/单克隆抗体/抗原表位Key words
African swine fever virus/D205R protein/Monoclonal antibody/Antigenic epitope分类
农业科技引用本文复制引用
林志钊,赵燕燕,任豪杰,史赛燕,韩世充,何文瑞,万博,张雨杭,张改平..非洲猪瘟病毒D205R蛋白单克隆抗体的制备和抗原表位鉴定[J].河南农业科学,2025,54(2):124-130,7.基金项目
国家自然科学基金项目(32272987,32102655,32373005) （32272987,32102655,32373005）
河南省重大科技专项(221100110600) （221100110600）
国家重点研发计划项目(2022YFD1801300) （2022YFD1801300）
河南省科技发展项目(222102110453) （222102110453）
