抗synuclein-γ单克隆抗体识别的抗原表位区域的确定  

Identification of antigenic epitope regions recognized by anti-synuclein-γ monoclonal antibodies

作　　者：谢玮[1] 苏亚辉[1] 刘彩云[1] 寿成超[1] 

机构地区：[1]北京大学临床肿瘤学院,北京肿瘤医院暨北京市肿瘤防治研究所生物化学与分子生物学研究室,恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142

出　　处：《北京大学学报（医学版）》2009年第3期361-367,共7页Journal of Peking University:Health Sciences

基　　金：国家高技术研究发展计划(“863计划”2006AA02A249);国家重点基础研究发展计划(“973计划”2009CB521800)项目~~

摘　　要：目的:确定已获得的多株抗突触蛋白γ(synulcein-γ,SNCG)单克隆抗体识别的抗原表位区域,为相关后续研究奠定基础。方法:分别构建谷胱甘肽疏基转移酶(GST)与SNCG不同截短体的表达载体并表达GST-SNCG截短体融合蛋白,采用ELISA和Western blot检测单抗与不同截短体融合蛋白的反应,推知不同单抗的抗原识别表位区域。依据推测的抗原识别表位的区域,构建并表达GST-表位小肽融合蛋白,并用相应单抗进行ELISA和Western blot检测,确定单抗的抗原表位区域。结果:确定了11株SNCG单抗的抗原识别表位区域,分别是9#、42#和3E4抗体的抗原表位位于SNCG第1到21位氨基酸,11#和22#抗体的抗原表位位于SNCG第78～92位氨基酸,1#、14#、18#、31#和36#抗体的抗原表位位于SNCG第93到110位氨基酸,6C8抗体的抗原表位位于SNCG第111～127位氨基酸。结论:确定了11株SNCG单抗的抗原识别表位区域,为SNCG的进一步研究和血清学检测奠定了基础。Objective：To identify the antigenic epitope regions recognized by anti-synuclein-γ mono- clonal antibodies. Methods： Six expression vectors of truncated SNCG were constructed and the trun- cated SNCG-GST fusion proteins were expressed in E. coli. The binding activities of anti-synuclein-γ （SNCG） monoclonal antibodies （mAbs） with truncated SNCG were analyzed by ELISA and Western blot. According to the speculative epitope regions, expression vectors of potential epitopes were construc- ted and the GST-epitope fusion proteins were expressed in E. coli. And the fusion proteins were analyzed by ELISA and Western blot with mAbs to confirm the epitope regions. Results： Epitope regions recog- nized by eleven anti-SNCG mAbs were identified. The epitopes recognized by 9#, 42#, and 3E4 were lo- cated in the region from 1st to 21st amino acids of SNCG, 11#, 22# in the region from 78th to 92nd amino acids, 1#, 14#, 18#, 31#, 36# in the region from 93rd to ll0th amino acids, and 6C8 in the region from l llth to 127th amino acids. Conclusion： The epitope regions recognized by eleven anti-SNCG mAbs were identified. It will benefit the research on SNCG and application of anti-SNCG monoclonal antibodies.

关 键 词：癌基因 抗体 单克隆 免疫显性表位 乳腺肿瘤 

分 类 号：R730.2[医药卫生—肿瘤]