南阳师范学院学报2025，Vol.24Issue(1)：46-52,7.
禽白血病P27蛋白单克隆抗体的制备
Monoclonal antibody preparation to Avian Leukosis P27 protein
摘要
Abstract
In order to improve the detection efficiency of Avian Leukosis Virus(ALV)in poultry farms,the P27 protein of Avian Leukosis Virus Subgroup J(ALV-J)was prepared by a prokaryotic expression system,and pre-pared a highly specific monoclonal antibody based on this protein in this study.Firstly,mouse ascites induced by hybridoma cell lines were obtained by immunological reaction with BABL/c mouse antigens.Subsequently,mon-oclonal antibodies were purified from ascites using octanoic acid-ammonium sulfate precipitation and Protein G agar gel affinity chromatography column method.The potency and specificity of the purified antibodies were iden-tified by Enzyme Linked Immunosorbent Assay(ELISA).Finally,the baculovirus expression system was used to express the P27 protein,and the specificity of the prepared antibody was tested.The results showed that four strains of hybridoma cells(3C9,1H15,1H6 and 7G2)were successfully obtained in this experiment.7G2,which was used as an enzyme-labelled antibody,showed the best performance,while 3C9 was the best antigen.The results by Indirect Immunofluorescence Assay(IFA)showed that the monoclonal antibodies were able to re-act specifically with baculovirus expressing avian leukaemia P27.This indicated that the monoclonal antibody to avian leukaemia P27 protein was successfully prepared.关键词
家禽养殖/禽白血病病毒/单克隆抗体/杆状病毒表达系统Key words
poultry farming/Avian Leukosis Virus/monoclonal antibodies/baculovirus expression system分类
生物工程引用本文复制引用
李洁琼,罗明展,王雄,胡小敏,张梦杨,姚伦广..禽白血病P27蛋白单克隆抗体的制备[J].南阳师范学院学报,2025,24(1):46-52,7.基金项目
河南省中原科技创新领军人才项目(244200510040) （244200510040）
河南省本科高校研究性教学改革研究与实践项目(2022SYJXLX079) （2022SYJXLX079）
