發(fā)布時(shí)間：2018-05-17 15:36

  本文選題：c-Met + 噬菌體抗體庫�。� 參考：《南京醫(yī)科大學(xué)學(xué)報(bào)(自然科學(xué)版)》2008年06期

【摘要】：目的:構(gòu)建大容量人源天然Fab噬菌體抗體庫,篩選抗c-Met特異性抗體并進(jìn)行初步鑒定。方法:采集20位健康成人的骨髓淋巴細(xì)胞,用PCR擴(kuò)增人Fab片斷抗體基因,插入載體pComb3XSS內(nèi),構(gòu)建人源天然Fab抗體庫。以固相化的抗原對抗體庫進(jìn)行6輪篩選后,隨機(jī)挑選60個克隆用Phage ELISA、BstOⅠ酶切片斷分析進(jìn)行檢測,陽性克隆作可溶性表達(dá)和鑒定。結(jié)果:構(gòu)建的Fab噬菌體抗體庫的庫容為1.2×109,從中篩選到1株與c-Met特異性結(jié)合的人源抗體克隆,命名為AM2-26。DNA序列分析證明為人免疫球蛋白可變區(qū)基因,Western blot證實(shí)為人源抗c-Met Fab抗體片段,ELISA特異性鑒定陽性。結(jié)論:構(gòu)建了大容量人源天然Fab抗體庫,從中獲得1株抗c-Met人源Fab抗體片段,有望為抗腫瘤藥物的研制提供新的候選分子。
[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，

本文編號：1901899