中国兽医科学2026，Vol.56Issue(8)：1049-1057,9.DOI:10.16656/j.issn.1673-4696.2026.0144
猪瘟病毒E2蛋白单克隆抗体的制备与鉴定
Preparation and identification of monoclonal antibodies to classical swine fever virus E2 protein
摘要
Abstract
To prepare monoclonal antibodies against the E2 protein of classical swine fever virus(CSFV),the recombinant plasmid pCAGGS-E2-His was transfected into human embryonic kidney 293T(HEK-293T)cells for eukaryotic expression and purification of the recombinant E2 protein,followed by immuniza-tion of BALB/c mice.Hybridoma cell lines stably secreting specific monoclonal antibodies against the E2 protein were screened using cell fusion technology.The reactivity of the monoclonal antibodies was verified by indirect immunofluorescence assay(IFA)and Western blot,and antibody subclass identifica-tion was performed after purification.The results showed that the recombinant E2 protein was success-fully expressed in HEK-293T cells,with a specific band of approximately 55 kDa detected.In this study,three hybridoma cell lines stably secreting anti-E2 monoclonal antibodies were obtained and designat-ed as 4B11E6,5E9G2 and 6B9F8,respectively.The titers of these monoclonal antibodies were above 1∶64000 with favorable specificity,which could recognize the purified CSFV E2 protein by Western blot and CSFV-infected porcine kidney(PK-15)cells by IFA.Antibody subtype analysis revealed that the heavy chains of 4B11E6 and 6B9F8 belong to the IgG1 subtype,while that of 5E9G2 belongs to the IgG2a subtype,and all light chains are of the κ isotype.In conclusion,the CSFV E2 protein was successfully expressed and high-titer monoclonal antibodies were prepared in this study,which provide important materials for further research on the function of the E2 protein and the establishment of diagnostic methods for classical swine fever.关键词
猪瘟病毒/E2蛋白/真核细胞表达/单克隆抗体Key words
classical swine fever virus/E2 protein/eukaryotic expression/monoclonal antibodies分类
农业科技引用本文复制引用
李璐瑶,孙国荣,郑海学,付强,郭庆勇,王琦,张宇,毛文权,吴世朋,刘阳河,段月月,袁聪,施磊,孙茂文..猪瘟病毒E2蛋白单克隆抗体的制备与鉴定[J].中国兽医科学,2026,56(8):1049-1057,9.基金项目
国家自然科学基金面上项目(32473022) （32473022）
中国农业科学院科技创新工程项目(CAAS-ASTIP-2021-LVRI) （CAAS-ASTIP-2021-LVRI）
中国农业科学院杰出青年英才项目 （）
动物疫病防控全国重点实验室人才培育项目(2025SKLADCP01) （2025SKLADCP01）
