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GST融合蛋白制备表皮生长因子受体抗体
引用本文:阎志勇,吴爱群,由振东,路长林,何成. GST融合蛋白制备表皮生长因子受体抗体[J]. 郑州大学学报(医学版), 2004, 39(2): 252-254
作者姓名:阎志勇  吴爱群  由振东  路长林  何成
作者单位:第二军医大学神经生物学教研室,上海,200433;郑州大学基础医学院人体解剖学教研室,郑州,450052;郑州大学基础医学院人体解剖学教研室,郑州,450052;第二军医大学神经生物学教研室,上海,200433
基金项目:国家自然科学基金资助项目 30070167,海外青年学者合作研究基金资助项目 30028003,国家“973”基金资助项目 G1999054000
摘    要:目的:探讨制备大量表皮生长因子受体(EGFR)特异性抗体的方法。方法:将编码EGFR的N端肽段的cDNA克隆入表达载体pGEX-4T-3中,转化大肠杆菌BL-21,经IPTG诱导表达后,经亲和层析柱纯化,用该GST融合蛋白免疫BALB/C小鼠,获得免疫血清,以ELISA和Western Blot检测抗体滴度。结果:获得GST—PTB融合蛋白纯度大于95%;经免疫小鼠抗血清滴度1:38827~1:190602,得到大量高滴度的EGFR特异性抗体。结论:用原核系统表达GST融合蛋白直接免疫小鼠可以快速、大量地制备EGFR特异性抗体。

关 键 词:EGFR  重组蛋白  抗体
修稿时间:2003-08-05

Preparation EGFR antibody with GST fusion protein
YAN Zhiyong,WU Aiqun,YOU Zhendong,LU Changlin,HE Cheng. Preparation EGFR antibody with GST fusion protein[J]. Journal of Zhengzhou University: Med Sci, 2004, 39(2): 252-254
Authors:YAN Zhiyong  WU Aiqun  YOU Zhendong  LU Changlin  HE Cheng
Affiliation:YAN Zhiyong,WU Aiqun,YOU Zhendong,LU Changlin,HE Cheng Department of Neurobiology,Second Military Medical University,Shanghai 200433 Department ofHuman Anatomy,Basic Medical College,Zhengzhou University,Zhengzhou 450052
Abstract:Aim: To produce the multi-antibody derived from mouse. Method: The 96bp cDNA encoding EGFR was amplified by using PCR method, and cloned into pGEX-4T-3 vector. The expression plasmid was introduced into E. coli strain BL-21. The GST-EGFRAg fusion protein was purified to homogeneity from the cell lysates via affinity after IPTG-in-duction. BALB/C mouse was immuned by using this purified fusion protein directly. Those antibody titrations were detected by ELISA and western blot. Result: High purified GST-EGFR fusion protein and high titration specified antibody was obtained. Conclusion : The established method for preparating large quantity EGFR antibody is effective.
Keywords:EGFR  recombinant protein  antibody
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