中国兽医科学2018，Vol.48Issue(9)：1102-1108,7.DOI:10.16656/j.issn.1673-4696.2018.0171
马立克病病毒单克隆抗体的制备
Development of monoclonal antibodies against Marek's disease virus
摘要
Abstract
Specific monoclonal antibodies against Marek's disease virus(MDV) were produced using the very virulent strain GX0101 as the immune antigen,which was purified from the virus-infected chicken embryo fibroblast(CEF) cultures. At 96 hours post-infection(hpi) ,the CEF cells were collected, frozen and thawed for 3 times. The supernatant was concentrated using a tangential flow ultrafiltration centrifuge tube and centrifuged and purified through the Sepharose 4 Fast Flow(4FF) gel as a chromatographic medium. Different collection peaks were further analyzed by polymerase chain reaction(PCR) and quantitative real-time PCR. MDV particles about 100 nm in diameter were observed by transmission electron microscope. The peaked virus particles concentrated by 100 ku tangential flow ultrafiltration centrifuge tube were used to immunize female BALB/c mice to prepare monoclonal antibodies. After cell fusion,a total of 107 positive hybridoma clones were finally confirmed by immunoperoxidase monolayer assay(IPMA) and Western-blot analysis. Clone 10B2,which was the strongest positive hybridoma cell line determined by IPMA(titer of 1 : 2. 56×106),was finally produced and characterized. The result provided an important basis for the future development of MDV monoclonal antibody associated diagnostic reagents.关键词
马立克病/马立克病病毒/单克隆抗体/IPMAKey words
Marek's disease/Marek's disease virus/monoclonal antibody/IPMA分类
农业科技引用本文复制引用
宋利娜,滕蔓,郑鹿平,李会珍,朱志坚,薛正飞,郅玉宝,罗俊,张改平..马立克病病毒单克隆抗体的制备[J].中国兽医科学,2018,48(9):1102-1108,7.基金项目
国家自然科学基金资助项目(U1604232,31602050) （U1604232,31602050）
国家重点研发计划项目(2016YFD0500800) （2016YFD0500800）
