中国免疫学杂志2011，Vol.27Issue(12)：1119-1122,1128,5.DOI:10.3969/j.issn.1000-484X.2011.12.16
抗人gp130单克隆抗体的制备及其生物学特性分析
Preparation of the monoclonal antibody against human gp130 and analysis for its biological traits
摘要
Abstract
0bJ3ctive :To prepare antrhum an gpl30 m onocbnal antibodies ,and prelin inary analysis of 1heir bbbgical characteristics and flmctiDns.M e1hods:lnmunized the BALB/cm ice with gpl30,1hen fusbn 1he spleen cells of 1heBALB/cm ice and 1heSP2/0 eels ,cul- lured in HAT screened m edium ,after several subebne ,harvested several hybridan a cell lines secreting antrgpl30 m onocbnal antibody .Collect-ed 1he supernatant for affinity purification, using W estem bbt,indirectELBA and biacore ti anafyze the purity, specificity and affinily of antibodies ,obseived 1he can binatin of antibody w ith U 266 cells by FACS ,and asessed 1he proliferation inhibitay effects on U 266 eel under condi- tfansof L~6 existence .Results:Severalanti hum an m onocbnal antibodies onfy against gpl30were harvested, se]ectone of these (14C4) for the folbw ing study ,the subiypes bebnged to ]gG2b ,k light chain ,W estem bbt and indirect ELBA dan onstrated that it had a higher purity and specificity,biacore showed that the binding affinily was 2 .62EH.0 ,FACS results showed that it couli specific bind the gpl30 to the surface of U266 cells and in a dose-dependentmanner,cellproliferatiDn test showed :mAb inhibited L~6 induced U266 cell proliferation .Conclusion :W e prelin inary prepared the antrgpl30 m onocbnal antibody,and it had a better bbbgicall characteristics,it supplied effective tools for the study of him an L-6gpl30 anti-infective, inflammatory and autoinmune diseases.关键词
IL-6/gp130/单克隆抗体/特异性/亲和力Key words
L~6 / gpl30 /M onocbnal antibody/ SpeciOcily /A fHnity分类
医药卫生引用本文复制引用
苗平,钱柳,赵蓉,余奇文,张继英,陈雪华,陆梅生,张冬青..抗人gp130单克隆抗体的制备及其生物学特性分析[J].中国免疫学杂志,2011,27(12):1119-1122,1128,5.基金项目
本文受上海市免疫学研究所项目(08-A04)、上海市科委重点项目(10JC1408500)、上海市科委重点学科(外科学)开放课题(S30204-K01)等基金资助 （08-A04）
