人组织激肽释放酶的生物信息学分析及其不同抗原表位的多克隆抗体的制备及纯化  

Bioinformatics analysis of tissue kallikrein and preparation of peptide-directed polyclonal antibody against multiple epitopes on surface of an antigen

作　　者：冉晓[1] 陈昊[2] 张琴[2] 

机构地区：[1]华中科技大学同济医学院附属同济医院急诊科,武汉430030 [2]华中科技大学同济医学院附属同济医院麻醉科,武汉430030

出　　处：《中国免疫学杂志》2014年第4期491-494,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金(81100864);华中科技大学技术创新基金(01-18-540099)资助

摘　　要：目的:利用生物信息学分析人组织激肽释放酶(TK),并制备针对TK不同抗原表位的多克隆抗体。方法:根据生物信息学分析TK的结果为依据,选取针对TK不同表位的2段氨基酸序列合成多肽并与血蓝蛋白(KLH)偶联。免疫家兔,获得多克隆抗体并纯化,分别用ELISA,SDS-PAGE染色及Western blot等方法进行滴度,纯度,特异性及抗原表位分析。结果:制备的抗TK多克隆抗体的纯度均为100%,其效价均在1∶25 600以上,具有较高的特异性并且识别不同的抗原表位。结论:利用生物信息学软件成功地预测出TK的抗原性,并制备了针对其不同表位的多克隆抗体。Objective：To analyze tissue kallikrein （TK） by bioinformatics and prepare its polyclonal antibody recognizing mul- tiple epitopes on the surface of an antigen. Methods ： According to the bioinformatics analysis and prediction of the possible high struc- ture, hydrophobicity and antigenicity of TK and the principal of antibody project, the synthetic TK peptide consisting of 11 and 12 amine acids （ 11P and 12P） were fused to keyhole limpet hemocyanin （KLH） and used to immunize rabbit. Antibody was then purified by af- finity column chromatography. The specificity and titer of polyclonal antibodies against TK were identified by ELISA and Western blot. SDS-PAGE was used to visualize the purified antibody. Results： Polyclonal antibodies against TK were proved to recognize different epitope on the surface of TK specifically, and its titers reached 1：25 600. Tile purity of polyclonal antibody was 100%. Conclusion： By the bioinformatics analysis and prediction, the hydrophobicity and antigenicity of TK are analyzed. The polyclonal antibody against TK recognizing multiple epitopes on the surface of an antigen has been successfully obtained.

关 键 词：人组织激肽释放酶 多肽 表位 多克隆抗体 纯化 

分 类 号：R392.33[医药卫生—免疫学]