畜牧与兽医2025，Vol.57Issue(6)：77-82,6.
猪A群轮状病毒VP4单克隆抗体的制备和初步鉴定
Preparation and preliminary characterization of monoclonal antibodies against VP4 of porcine rotavirus A
摘要
Abstract
This study aimed to prepare a monoclonal antibody against the VP4 protein of porcine rotavirus A(PoRVA)with strong specific-ity and high sensitivity.In here,the VP4 gene(2 463 bp)of PoRVA was amplified by PCR,and prokaryotic expression vector pET-28a-VP4 was constructed to express the protein.Finally,monoclonal antibodies were prepared to identify it.The results showed that the recombi-nant protein was about 98 kDa,was insoluble and existed in the inclusion bodies.The His-tagged antibody and PoRV positive serum could recognize the recombinant protein.The band was single and clear,and the protein had good antigenicity.After protein purification,the mouse immune cell fusion filtering process,and three rounds of cloning,a positive hybridoma cell line 4B10 was finally obtained which was able to steadily produce monoclonal antibodies against VP4 proteins.The light chain type of the strain was Kappa,its heavy chain subtype was of IgG1,and its titer of mouse ascites preparation was 1∶3 000 000.On this basis,the monoclonal antibody prepared was able to specifically recognize PoRV Pig/NJ/2012 with good reactivity,and can be used for direct detection of target antigens;which provided a biological mate-rial for subsequent application of monoclonal antibodies.In conclusion,the monoclonal antibody prepared in this study had strong specificity and high sensitivity,providing biological materials for future research on the protein function and the pathogenic mechanism of the virus.关键词
猪A群轮状病毒/单克隆抗体/VP4蛋白Key words
porcine rotavirus A/monoclonal antibody/VP4 protein分类
农业科技引用本文复制引用
毛玎懿,孙波涛,戴琦,李文良,李梅珍,陈艳,赵冰倩,陈婧,周斌..猪A群轮状病毒VP4单克隆抗体的制备和初步鉴定[J].畜牧与兽医,2025,57(6):77-82,6.基金项目
国家重点研发计划项目(2021YFD1801300) （2021YFD1801300）
