抗GSTAl单克隆抗体的制备与鉴定

Preparation and Identification of the Monoclonal Antibody Against Humam

目的:制备鼠抗人谷胱甘肽S转移酶A1(GSTAI)的单克隆抗体(mAb),并进行特性鉴定。方法:用纯化后的GSTAl融合蛋白作为抗原免疫BALB/c小鼠,采用常规的细胞融合技术制备抗GSTAl的单克隆抗体(mAb)。用ELISA法测定鼠抗人GSTA1血清的效价;用Western blot方法鉴定抗血清的特异性。结果:筛选到l株可稳定分泌抗人GSTAlmAb的细胞株。腹水效价为12 800。Western blot显示在相对分子量(Mr)约25KD处出现特异性条带。说明制备的mAb可特异地识别成人肝细胞中相对分子量(Mr)约25KD的GST-GSTA1。结论:获得一株能特异性识别抗人GSTAI的单克隆抗体,可用于肝损伤性疾病的诊断。

Objective：To prepare the mouse monoclonal antibodies against human GSTA1 and characterize its properties. Methods： Used GST - GSTA1 as the immunogen to prepare the mouse monoclonal antibodies. The properties of antiserum against human GSTA1 were characterized by ELISA and Western blotting. Results： Acquired one done of cell that can excrete monoclonal antibodies against human GSTA1 stably. The GST - GS- TA1 fusion protein was highly expressed with a molecular weight of 25 KDa, The titre of ascites against human GSTA1 was 1：12 800. Conclusions： The successful generation monoclonal antibodies the GST- GSTA1 fusion protein will provide efficient affinity reagents for the further functional studies of GSTA1 expressed in human hepatocellular impairment.