磷脂酰肌醇蛋白聚糖3的原核表达及单克隆抗体制备

生物技术通报 ›› 2014, Vol. 0 ›› Issue (3): 177-181.

磷脂酰肌醇蛋白聚糖3的原核表达及单克隆抗体制备

程华¹, 方向东², 吴萌¹, 史蕾³, 崔硕¹

(1. 河北省科学院生物研究所，石家庄 050081；2. 中国科学院北京基因组研究所，北京 100101；3. 河北师范大学生命科学学院，石家庄 050000)

收稿日期:2013-08-29 出版日期:2014-03-29 发布日期:2014-03-31
作者简介:程华，男，副研究员，硕士，研究方向:肿瘤早期诊断；E-mail:cheng_hua@sina.com
基金资助:
河北省科学院科技攻关项目(13335)

Expression and Antibody Preparation of Glypican3

Cheng Hua¹, Fang Xiangdong², Wu Meng¹, Shi Lei³, Cui Shuo¹

(1. Hebei Institue of Biology，Shijiazhuang 050081；2. Beijing Institute of Genomics，Chinese Academy of Sciences，Beijing 100101；3. College of Life Science，Hebei Normal University，Shijiazhuang 050000)

Received:2013-08-29 Published:2014-03-29 Online:2014-03-31

摘要/Abstract

摘要： 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。

关键词: 磷脂酰肌醇蛋白聚糖3, 蛋白表达, 单克隆抗体

Abstract: 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.

Key words: Glypican-3, Protein expression, Monoclonal antibody

程华, 方向东, 吴萌, 史蕾, 崔硕. 磷脂酰肌醇蛋白聚糖3的原核表达及单克隆抗体制备[J]. 生物技术通报, 2014, 0(3): 177-181.

Cheng Hua, Fang Xiangdong, Wu Meng, Shi Lei, Cui Shuo. Expression and Antibody Preparation of Glypican3[J]. Biotechnology Bulletin, 2014, 0(3): 177-181.

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