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.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
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)~~
关键词
Ⅰ型丙氨酸氨基转移酶
单克隆抗体
制备与鉴定
I-alanine aminotransferase
monoclonal antibody
preparation
identification
作者简介
李有强，男，湖南省娄底市人，硕士，检验师，主要从事临床免疫学诊断方面的研究。E-mail：liyouqiang21@126．com
通信作者：尹一兵，电话：（023）68485658，E-mail：yibingyin56@126.com
