人Ⅰ型丙氨酸氨基转移酶单克隆抗体的制备及鉴定  

Preparation and identification of monoclonal antibody against human I-alanine aminotransferase

作　　者：李有强[1] 尹一兵[1] 胥文春[1] 张云燕[2] 张雪梅[1] 尹楠林[1] 

机构地区：[1]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学附属口腔医院牙体牙髓科,重庆400010

出　　处：《第三军医大学学报》2009年第20期2001-2004,共4页Journal of Third Military Medical University

基　　金：重庆市教委科技项目(000106)~~

摘　　要：目的通过构建人Ⅰ型丙氨酸氨基转移酶(ALT1)的原核表达载体,并用纯化的ALT1重组蛋白免疫BALB/c小鼠,制备有生物活性的单克隆抗体。方法用GST-ALT1融合蛋白作为免疫原,通过杂交瘤技术制备抗ALT1的单克隆抗体,用间接ELISA、Dot-Elisa、Westernblot等方法对抗体的抗原结合特性进行鉴定。结果获得1株能稳定分泌抗ALT1单克隆抗体的细胞株8A9,亚类鉴定这种单抗为IgG2。间接ELISA法测定细胞株腹水抗体的效价为10×105。Dot-ELISA、Western blot显示ALT1单克隆抗体能特异性地识别免疫原。结论成功制备ALT1单克隆抗体。Objective To prepare the bioactive monoclonal antibody （McAb） against the fusion protein of GST-human I-alanine aminotransferase （ALT1）. Methods With the fusion protein GST-ALT1 as immunogen, the anti -ALT1 McAb was produced by using the hybridoma technique, of which the antigen binding characteristics was identified by indirect enzyme-linked immunosorbent assay （ELISA）, Dot-ELISA and Western blot assay.Results One cell strain of hybridoma was obtained and named as 8A9 which secreted the anti-ALT1 McAb. The class and subtype identification demonstrated that this cell strain was IgG2.The titers of ascites fluid which hybridoma induced was 10×105. Dot-ELISA and Western blotting demonstrated that this antibody recognized specifically the immunogen and ALT1 which existed in clinical serum specimen. Conclusion McAb against ALT1 is prepared successfully and provides a useful tool for researching on ALT1’s property, function, bionomics and found a basis for establishment of specific immunoassays of human ALT1.

关 键 词：Ⅰ型丙氨酸氨基转移酶 单克隆抗体 制备与鉴定 

分 类 号：R345.5[医药卫生—基础医学] R392.33