人源天然Fab噬菌体抗体库的构建及抗c-Met抗体的筛

发布时间：2018-05-17 15:36

  本文选题：c-Met + 噬菌体抗体库　； 参考：《南京医科大学学报(自然科学版)》2008年06期

【摘要】：目的:构建大容量人源天然Fab噬菌体抗体库,筛选抗c-Met特异性抗体并进行初步鉴定。方法:采集20位健康成人的骨髓淋巴细胞,用PCR扩增人Fab片断抗体基因,插入载体pComb3XSS内,构建人源天然Fab抗体库。以固相化的抗原对抗体库进行6轮筛选后,随机挑选60个克隆用Phage ELISA、BstOⅠ酶切片断分析进行检测,阳性克隆作可溶性表达和鉴定。结果:构建的Fab噬菌体抗体库的库容为1.2×109,从中筛选到1株与c-Met特异性结合的人源抗体克隆,命名为AM2-26。DNA序列分析证明为人免疫球蛋白可变区基因,Western blot证实为人源抗c-Met Fab抗体片段,ELISA特异性鉴定阳性。结论:构建了大容量人源天然Fab抗体库,从中获得1株抗c-Met人源Fab抗体片段,有望为抗肿瘤药物的研制提供新的候选分子。
[Abstract]:Aim: to construct a large human natural Fab phage antibody library and to screen and identify anti c-Met specific antibodies. Methods: human Fab fragment antibody gene was amplified by PCR from bone marrow lymphocytes of 20 healthy adults and inserted into vector pComb3XSS to construct human natural Fab antibody library. After six rounds of screening the antibody library with solid antigen, 60 clones were randomly selected and detected by Phage ELISAN BSTO 鈪，

本文编号：1901899