新的人突触相关蛋白(FRG4)抗原表位分析及抗体制备  

Epitope analysis of a novel Homo spains synapse associated protein(FRG4) and preparation of antibody

作　　者：王仁[1] 杨向东[1] 屈顺林[1] 刘俊文[1] 闾宏伟[1] 姬志娟[2] 盛树力[2] 

机构地区：[1]南华大学分子生物中心,衡阳421001 [2]首都医科大学北京宣武医院多肽合成室,北京100053

出　　处：《中国免疫学杂志》2005年第7期531-534,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金(30200103);湖南省自然科学基金(02JJY4011)资助

摘　　要：目的:分析一个新的人突触相关蛋白(FRG4)抗原表位并制备其多克隆抗体。方法:从人胎肝文库PCR扩增获得FRG4基因全长cDNA序列;通过生物信息学分析,预测FRG4编码氨基酸序列的二级结构、抗原决定簇、功能结构域,并进行了多序列比对;采用固相多肽合成法合成了FRG4抗原多肽,并免疫家兔;用免疫组化检测蛋白在人肝癌细胞HepG2细胞中的表达。结果:通过生物信息学分析选取抗原13肽PKLVKEEVFWRNY,制备兔抗人FRG4多克隆抗体。高效液相色谱检测显示抗体纯度达82.79%,抗体滴度为1∶16000,Westernblot证实该抗体具有较好的反应性和特异性,免疫组化证实其主要在HepG2细胞胞浆中表达。结论:成功制备了新的人突触相关蛋白(FRG4)多克隆抗体。Objective:To analyze a novel epitope of Homo sapiens synapse associated protein and synthesize polyclonal antibody.Methods:FRG4 full-length sequence was obtained by PCR from human fetal liver library;by bioinformatics to detect the second structure of amino acids encoded by FRG4 and its epitope and motifs;by solid-phase peptide synthesis method to synthesize FRG4 peptides,then peptides were immunized to rabbits;by immunohistory to detect the expression of FRG4 in HepG_2 cells.Results:Select 13-peptides PKLVKEEVFWRNY by bioinformatics to synthesize rabbit anti-human FRG4 polyclonal antibody.Antibody purity was 82.79% and antibody dilution was 1∶16 000 detected by ELISA.The antibody had a good reaction and speciality in Western blot,it was mainly expressed in cytoplasm of HepG_2 cells.Conclusion:A novel Homo sapiens synapse associated protein(FRG4) antibody was synthesized successfully.

关 键 词：巨噬细胞 动脉粥样硬化 突触相关蛋白 多肽合成 抗体 

分 类 号：R392.11[医药卫生—免疫学]