中国兽医科学2026，Vol.56Issue(4)：512-519,8.DOI:10.16656/j.issn.1673-4696.2026.0069
非洲猪瘟病毒K205R蛋白单克隆抗体的制备及功能鉴定
Preparation and characterization of monoclonal antibodies against African swine fever virus K205R protein
摘要
Abstract
To generate monoclonal antibodies(mAbs)against the K205R protein of African swine fever virus(ASFV)and characterize their biological properties,cell fusion technology was employed.The specificity of the antibodies was assessed using Western-blot,indirect immunofluorescence assay(IFA)and co-immunoprecipitation(Co-IP).Antibody titers and subtypes were determined via ELISA method,and virus neutralization activity was evaluated through neutralization assay.Results showed that the eukaryotic-expressed K205R protein had a molecular weight of approximately 23 kDa,consistent with the expected size.Five hybridoma cell lines secreting K205R mAbs were screened.Western-blot analysis demonstrated that all five mAbs specifically recognized the K205R protein in ASFV-infected cell lysates,without cross-reaction with samples infected with foot-and-mouth disease virus.IFA further confirmed their specific binding to K205R in both transfected and ASFV-infected cells.The highest an-tibody titer could reach 1∶819 200 as determined by indirect ELISA.Isotype analysis showed that all five mAbs belonged to the IgG1 subclass with kappa light chains.Neutralization assay revealed that one of the mAbs,A11-E11,exhibited certain neutralizing activity for ASFV.Co-IP assay also verified A11-E11 mAbs can bind K205R protein in ASFV-infected porcine alveolar macrophages.In conclusion,this study successfully produced monoclonal antibodies against the ASFV K205R protein with high specificity,pro-viding a foundation for further investigation into the biological function of K205R and valuable mate-rial for the development of diagnostic techniques for African swine fever.关键词
非洲猪瘟病毒/K205R蛋白/单克隆抗体/细胞融合技术Key words
African swine fever virus/K205R protein/monoclonal antibody/cell fusion technology分类
农业科技引用本文复制引用
叶银波,周斌,赵倩,黄丹妮,杨洋,孙波涛,程颜,刘永杰,秦晓东,郑海学..非洲猪瘟病毒K205R蛋白单克隆抗体的制备及功能鉴定[J].中国兽医科学,2026,56(4):512-519,8.基金项目
江苏省农业科技自主创新资金项目(CX(24)1010) （CX(24）
甘肃省联合科研基金重大项目(24JRRA806) （24JRRA806）
