抗HBV G145R HBsAg多克隆抗体的制备与鉴定  

The preparation and identification of anti-HBV G145R HBsAg polyclonal antibody

作　　者：李佩珊[1] 龚劲松[2] 李时君[1] 张波[2] 陈妍[2] 项美娟[1] 李方和[2] 

机构地区：[1]中国生物技术集团总公司武汉生物制品研究所,武汉430060 [2]华中科技大学同济医学院附属同济医院

出　　处：《中国实用医药》2008年第36期1-3,共3页China Practical Medicine

基　　金：国家973计划项目资助(项目编号:2005CB522901)

摘　　要：目的实验性rG145R变异抗HBs多克隆抗体。方法采用纯化全基因重组G145R变异HBsAg常规皮下免疫制备小鼠多克隆抗体,以间接ELISA、SDS-PAG及Western blot等多种实验对制备物进行鉴定,并初步应用于转染CHO细胞的化学染色。结果制备物在Western blot、ELISA中和试验与抗原替代ELISA中与重组G145R变异HBsAg及重组野生HBsAg等均有很好的特异性与反应性;对此两种抗原的ELISA效价前者显著高于后者(分别为51200与6400);将其用于2A8细胞(转染并分泌G145R HBsAg)爬片的PAP染色,亦取得较理想的实验结果。结论采用重组G145R变异HBsAg免疫成功制备出较高效价的抗体。鉴于该抗体同时与野生重组wHBsAg存在较强交叉反应,其与野生抗HBs的混合应用能增ELISA免疫逃逸变异的检测能力。Objective To prepate the mouse polyclonal Anti G145R rHBsAb and analyze its immuno- logical characteristics. Methods The recombinant G145R mutant HBsAg were respectively used to develop poly-clonal antibodys through immunize BALB/c rats, and the polyclonal antibody were confirmed by empirical methods, include western blot, ELISA and SDS PAGE, and preliminary used in chemical staining for the CHO. Results The recombinant Gl＇45R mutant HbsAb poly were indentified by western blot and ELISA, it showed a good specific reaction performence to both of G145R rHBsAg and r-wHBsAg. And the potency of the reaction with G145R rHBsAg was obviously higher than r-wHBsAg（ they were 51200 and 6400）. And it had a good result of the PAP staining of cell 2A8 also. Conclusion The satisfactory antibody could be created by immunizing with G145R rHBsAg. Because of the cross reaction with r-wHBsAg, the recombinant G145R mutant HbsAb poly which is mixed with r-wHBsAg can enhance the detectivity of ELISA.

关 键 词：乙型肝炎病毒 基因变异 合成肽 多克隆抗体 

分 类 号：R392.1[医药卫生—免疫学]