磷脂酰肌醇蛋白聚糖3的原核表达及单克隆抗体制备

Expression and Antibody Preparation of Glypican 3

人磷脂酰肌醇蛋白聚糖3(GPC3)是一个肝癌潜在的诊断标志物和治疗靶点。为获得抗GPC3单克隆抗体,用PCR方法克隆GPC3基因,利用酶切位点将该序列插入pGEX-2T载体,构建了pGEX-2T-gpc3表达载体。转化大肠杆菌BL21(DE3)菌株,经IPTG诱导,获得高效表达。经纯化后获得分子量为58 kD的蛋白。以该重组蛋白为免疫原,通过免疫BALB/c小鼠,经过细胞融合和筛选获得1株抗该重组蛋白的单克隆抗体1F3。Western blot分析显示,该抗体具有较好的特性,可用于GPC3蛋白的功能和应用研究。

Human glypican3 protein is a potential cancer therapeutic targets and diagnostic markers. In order to obtain anti-GPC3 monoclonal antibody, GPC3 gene was cloned into an expression vector pGEX-2T, which was transformed into E. coli BL21（DE3）. Induced by IPTG, a molecular weight of 58 kD protein was obtained. After immunizing BALB/c mice, cell fusion and screening, one monoclonal antibody was generated. Western blot analysis revealed that the antibody has a strong specificity and high titer can be used for GPC3 protein function and applied research.