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| 抗木瓜凝乳蛋白酶单链抗体基因的构建、初步筛选和鉴定 | |
| 引用本文： | 王舰平,赵树进,王捷,武婕,杨太成,杨静.抗木瓜凝乳蛋白酶单链抗体基因的构建、初步筛选和鉴定[J].细胞与分子免疫学杂志,2004,20(6):689-692. |
| 作者姓名： | 王舰平 赵树进 王捷 武婕 杨太成 杨静 |
| 作者单位： | 1. 华南理工大学生物工程系,广东,广州,510640 2. 广州军区广州总医院药学部,广东,广州,510010 3. 广州军区肿瘤分子生物学研究所,广东,广州,510010 |
| 基金项目： | 军队总后勤部科研基金资助项目 (No .0 1Q0 2 2 ) |
| 摘 要： | 目的 :构建抗木瓜凝乳蛋白酶全套单链抗体 (scFv)噬菌体表面展示文库。方法 :从木瓜凝乳蛋白酶免疫小鼠的淋巴细胞中提取总RNA ,反转录成cDNA后 ,用抗体V区简并引物扩增全套VH 和VL 基因。经重叠延伸反应 ,装配成scFv基因 ,并将其克隆到噬粒载体pFAB5C中 ,构建scFv噬菌体抗体库。对抗体库进行亲和筛选后 ,用ELISA法鉴定抗木瓜凝乳蛋白酶的scFv。结果 :经 4轮“亲和 吸附 洗脱”的富集过程 ,得到ELISA活性较高可与木瓜凝乳蛋白酶结合的克隆。结论 :成功地构建了抗木瓜凝乳蛋白酶的scFv噬菌体展示文库 ,并筛选到与木瓜凝乳蛋白酶具有结合能力的scFv基因 |
| 关 键 词： | 木瓜凝乳蛋白酶 可变区基因 单链抗体 噬菌体抗体库 |
| 文章编号： | 1007-8738(2004)06-0689-04 |
| 修稿时间： | 2004年3月15日 |
Construction and identification of anti-chymopapain scFv phage display library |
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| WANG Jian-ping,ZHAO Shu-jin,WANG Jie,WU Jie,YANG Tai-cheng,YANG Jing.Construction and identification of anti-chymopapain scFv phage display library[J].Journal of Cellular and Molecular Immunology,2004,20(6):689-692. | |
| Authors: | WANG Jian-ping ZHAO Shu-jin WANG Jie WU Jie YANG Tai-cheng YANG Jing |
| Institution: | Department of Bioengineering, South China University of Technology, Guangzhou 510640, China. bestwjp@sohu.com |
| Abstract: | AIM: To construct phage display library of anti-chymopapain scFv. METHODS: V_H and V_L gene repertoires were amplified from splenocyte mRNA by RT-PCR and joined by a (Gly_4ser)_3 linker to obtain scFv genes. The scFv genes were then cloned into phagemid pFAB5C to construct phage display library. Affinity selection and ELISA were used to (identify) specific phage antibody to chymopapain. RESULTS: After 4 rounds of panning, high affinity scFv was obtained. CONCLUSION: Phage display library of anti-chymopapain scFv was successfully constructed, and scFv with binding (ability) to chymopapain was obtained. |
| Keywords: | chymopapain scFv phage display library |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |