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基于B细胞表位制备抗肝细胞生成素的抗体
引用本文:周游,吴峰,范国才,王清明.基于B细胞表位制备抗肝细胞生成素的抗体[J].生物技术通讯,2009,20(6):761-764.
作者姓名:周游  吴峰  范国才  王清明
作者单位: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
基金项目:国家重大科学研究计划 
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。

关 键 词:肝细胞生成素  B细胞表位  噬菌体展示  抗体

Generation of Antibody Against Hepatopoietin Based on B Cell Epitopes
ZHOU You,WU Feng,FAN Guo-Cai,WANG Qing-Ming.Generation of Antibody Against Hepatopoietin Based on B Cell Epitopes[J].Letters in Biotechnology,2009,20(6):761-764.
Authors:ZHOU You  WU Feng  FAN Guo-Cai  WANG Qing-Ming
Institution:( State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China)
Abstract: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.
Keywords:hepatopoietin  B cell epitope  phage display  antibody
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