基于B细胞表位制备抗肝细胞生成素的抗体  

Generation of Antibody Against Hepatopoietin Based on B Cell Epitopes

作　　者：周游[1] 吴峰[1] 范国才[1] 王清明[1] 

机构地区：[1]蛋白质组学国家重点实验室,北京蛋白质组研究中心,军事医学科学院放射与辐射医学研究所,北京102206

出　　处：《生物技术通讯》2009年第6期761-764,共4页Letters in Biotechnology

基　　金：国家重大科学研究计划(2006CB910803)

摘　　要：目的:基于B细胞表位制备抗肝细胞生成素(HPO)的抗体。方法:根据HPO的空间结构选择了2个候选B细胞表位,展示在T7噬菌体的表面,将提取的重组噬菌体免疫动物,采用ELISA法检测抗血清的效价,通过杂交瘤技术制备针对HPOC端表位的单克隆抗体。结果:2个候选B细胞表位KDGSCD和DGWKDGSC均能诱导抗相应表位多肽的多克隆抗体的产生,免疫6周后血清中抗体效价均达到1∶103,产生的抗体还能够特异识别HPO全蛋白;针对HPOC端表位KDGSCD的单克隆抗体也能识别HPO全蛋白,且具有良好的特异性。结论:基于T7噬菌体展示的B细胞表位可作为免疫原用于制备识别该B细胞表位来源的全蛋白质的抗体。Objective： To make antibody against hepatopoietin（HPO） based on B cell epitopes. Methods： Based on the structure of HPO, two epitope peptides were selected and displayed on T7 phage. The recombinant phages were extracted and used to immunize animals. The titers of anti-sera antibody and their abilities to recognize HPO protein were moni- tored by ELISA. Hybridoma technology was used to generate monoclonal antibody against C-terminal epitope of HPO. Re- suits： The two alternative B cell epitopes, KDGSCD and DGWKDGSC, both induced the generation of antibodies against them. Six weeks later, titers of antiserum both reached 1：103. The antibodies could also recognize HPO protein. The mono- clonal antibody against C-terminal epitope of HPO could recognize HPO protein, and had good specificity. Conclusion： B cell epitope displayed on T7 phage could be used as immunogen to generate antibody against the source native protein.

关 键 词：肝细胞生成素 B细胞表位 噬菌体展示 抗体 

分 类 号：R392.1[医药卫生—免疫学] Q511[医药卫生—基础医学]