| GST融合蛋白制备表皮生长因子受体抗体 |
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| 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 |
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| 作者姓名： | 阎志勇 吴爱群 由振东 路长林 何成 |
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| 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 |
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| 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 |
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| 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。
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| 关 键 词： | EGFR 重组蛋白 抗体 |
| 修稿时间： | 2003-08-05 |
Preparation EGFR antibody with GST fusion protein |
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| 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 |
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| Authors: | YAN Zhiyong WU Aiqun YOU Zhendong LU Changlin HE Cheng |
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| 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 |
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| 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. |
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| Keywords: | EGFR recombinant protein antibody |
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