| 人源性抗角蛋白单链抗体的构建及表达 |
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| 引用本文： | 张国民,陈宇萍,王刚,刘玉峰.人源性抗角蛋白单链抗体的构建及表达[J].细胞与分子免疫学杂志,2005,21(6):719-722. |
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| 作者姓名： | 张国民 陈宇萍 王刚 刘玉峰 |
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| 作者单位： | 1. 首都医科大学微生物学和免疫学教研室,北京,100069;第四军医大学西京医院皮肤科,陕西,西安,710032
2. 第四军医大学西京医院皮肤科,陕西,西安,710032 |
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| 基金项目： | 国家高技术研究发展计划(863)资助(No.2001AA215361) |
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| 摘 要： | 目的:克隆抗角蛋白抗体的可变区(V区)基因,构建抗角蛋白单链抗体(scFv)表达载体并表达。方法:根据人源性抗角蛋白Fab段的基因序列设计引物,用PCR方法克隆抗角蛋白抗体的V区基因,并重组到scFv的表达载体中,在大肠杆菌进行表达,表达产物经体外变性复性后进行SDS-PAGE鉴定。用ELISA检测其特异性结合活性。结果:成功地克隆抗角蛋白抗体的V区基因,构建了scFv的表达载体,并在大肠杆菌中表达。ELISA检测证实,表达的scFv保留了亲本Fab抗体的特异结合活性。结论:获得功能性抗角蛋白的scFv,可用于进一步的临床应用研究。
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| 关 键 词： | 角蛋白 V区基因 scFv |
| 文章编号： | 1007-8738（2005）06-0719-04 |
| 收稿时间： | 2004-10-28 |
| 修稿时间： | 2005-05-25 |
Construction and expression of anti-keratin single-chain Fv antibody |
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| ZHANG Guo-min,CHEN Yu-ping,WANG Gang,LIU Yu-feng.Construction and expression of anti-keratin single-chain Fv antibody[J].Journal of Cellular and Molecular Immunology,2005,21(6):719-722. |
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| Authors: | ZHANG Guo-min CHEN Yu-ping WANG Gang LIU Yu-feng |
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| Institution: | Department of Microbiology and Immunology, Capital University of Medical Science, Beijing 100054, China. erminzhang@sohu.com |
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| Abstract: | AIM: To construct an expression vector of human anti-keratin scFv and express functional scFv in (E.coli). METHODS: The V_H and V_L gene were amplified by PCR and cloned into a scFv expression vector, which was expressed in E.coli. The expression of scFv was tested by SDS-PAGE; The antigen-binding characteristics of scFv were tested by ELISA. RESULTS: The V_H and V_L gene were cloned and expressed in E.coli successfully. The expressed scFv retained the antigen-binding characteristics (of the) parental antibody. CONCLUSION: A functional anti-(keratin) scFv was obtained. |
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| Keywords: | keratin V-gene scFv |
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