鼠伤寒沙门菌延伸因子EF-Tu蛋白单克隆抗体的制备及鉴定  

Preparation and identification of the monoclonal antibody against elongation factor Tu of Salmonella typhimurium

作　　者：张怡聪[1,2] 张杰[1,2] 房健[1,2] 吕俊峰[1,2] 刘思当[1,2] 赵孝民[1,2] 王方昆[1,2] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]山东省动物生物工程与疾病防治重点实验室,山东泰安271018

出　　处：《中国兽医科学》2015年第2期137-141,共5页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31201929)

摘　　要：为制备鼠伤寒沙门菌延伸因子Tu(EF-Tu)的单克隆抗体,以原核表达并纯化的鼠伤寒沙门菌EF-Tu蛋白为免疫原,免疫BALB/c小鼠,分离脾细胞与SP2/0融合,以间接ELISA筛选出阳性克隆,经5次亚克隆后获得5株稳定分泌抗EF-Tu蛋白抗体的杂交瘤细胞株,遂将其命名为Tu1-A1、Tu1-C4、Tu1-H2、Tu4-C11、Tu9-B9。对这5株单克隆抗体的初步鉴定结果表明,其抗体亚型均为IgG1/κ型。以原核表达的带GST标签的EF-Tu蛋白做Western-blot检测制备的单克隆抗体,证实所制备的单克隆抗体具有良好的反应性。这些单克隆抗体的制备为后期研究EF-Tu蛋白在沙门菌侵入及胞内存活过程中的调控作用奠定了基础。To prepare the monoclonal antibody(McAb)against elongation factor Tu(EF-Tu)protein of Salmonella typhimurium,six-week-old female BALB/c mice were immunized with purified recombinant ET-Tu of S.typhimurium.Five hybridoma cell lines stably secreting McAbs against EF-Tu protein were identified by an indirect ELISA.The subtypes of McAbs were identified as IgG1 subclass withκlight chain.Western-blot analysis showed that McAbs positively reacted with EF-Tu protein of S.typhimurium.The McAbs against EF-Tu protein developed in the present study would serve as useful tool to investigate the pathogenic processes of Salmonella bacterial invasion and survival within host cells.

关 键 词：鼠伤寒沙门菌 EF-Tu蛋白 GST标签 BALB/C小鼠 单克隆抗体 

分 类 号：S852.61[农业科学—基础兽医学]