福建农业学报2017，Vol.32Issue(8)：818-822,5.DOI:10.19303/j.issn.1008-0384.2017.08.002
猪流行性腹泻病毒单克隆抗体的制备及特性分析
Preparation and Characterization of Monoclonal Antibodies Against Porcine Epidemic Diarrhea Virus
摘要
Abstract
This study aimed to obtain the monoclonal antibodies against porcine epidemic diarrhea virus(PEDV) for antibody characterization as well asdisease control.The antigen of PEDV was purifiedby means of differential centrifugation and sucrose gradientcentrifugation.Spleen cells of the Balb/c mice immunized with viral antigen were fused with SP2/0 myeloma cells.Hybridoma cell lines secreting the monoclonal antibodies were isolated by using the indirect ELISA and sub-cloning approach to provide the source for the target antibody.Subsequently,specificities of the antibody were determined by means of IFA,ELISA and the Western blotting.The results obtained the following.(a) Two isolated hybridoma cell lines that could steadily secretethe specific McAbs against PEDV were designated as E1 and H6.(b) The isotyping analysis showed that E1 belonged to IgG2a;and H6,IgM sub-type.The McAbs from them could both bind to the antigen of PED in ELISA,IFA and the western blot analyses.(c) ELISA titers of the supernatants of E1 and H6 were 26 and 24 respectively;while those of the ascites fluids of E1 and H6,105 and 104 respectively.(d) The McAbs from E1 and H6 showed no cross-reactivity with Veto cells,CSFV,TGEV or PRRSV.(e) The Western blotting confirmed that E1 McAbs recognized M protein;and,H6 McAbs,N protein.It appeared that two monoclonal antibodies with a high specificity against PEDV were successfully prepared,and further studies on PEDV immunodiagnosis,epitope discernment and the proteins became possible.关键词
猪流行性腹泻病毒/单克隆抗体/制备/特性Key words
porcine epidemic diarrhea virus/monoclonal antibody/preparation/characterization分类
农业科技引用本文复制引用
王隆柏,王晨燕,吴学敏,车勇良,陈如敬,周伦江..猪流行性腹泻病毒单克隆抗体的制备及特性分析[J].福建农业学报,2017,32(8):818-822,5.基金项目
福建省自然科学基金项目(2015J01112) （2015J01112）
福建省科技创新平台建设项目(2014N2003) （2014N2003）
福建省科技计划项目——省属公益类科研院所基本科研专项(2014R1023-7) （2014R1023-7）
