人抗E．Coli J5噬菌体抗体制备的初步研究

以E．Coli J5株对合人Ig基因的噬菌体抗体库进行淘筛富集，免疫印迹筛选，以及ELISA检测，结果获得4株能与E．Coli J5株结合的阳性克隆，且阳性克隆结合抗原的活性可分别被E．Coli J5株、E．Coli Rc-LPS和抗E．Coli J5株核心糖脂域MAb抑制．PCR检测表明，4株阳性克隆均分别带有约660bp大小的重链和轻链基因片段．SDS－PAGE与蛋白质印迹的结果显示，经IPTG诱导的阳性克隆能表达分子量约为50000大小的蛋白，提示该4株阳性克隆能够表达具有一定抗原结合活性的人源Fab片段．

Preliminary Study on Preparation of Human Phage Antibodies to E. coli J5 Strain

The clones of phage antibody (PhAb)to E. coli J5 strain were enriched and screened by panning and filter blot with E. coli J5 strain from a human antibody library. Four positive clones binding to E. coli J5 strain were determined by ELISA. The results of inhibition test showed that the binding of positive clones to antigens could be respectively inhibited by E. coli J5 strain, E. coli Rc-LPS and anti-CGL MAbs to E. coli J5 strain. PCR amplification showed that all of the four positive clones contained the expected heavy and light chain genes of human Fab fragment. SDS-PAGE and Western blot analysis confirmed that positive clones were able to express about 50 ku proteins after induced with IPTG. These data indicate that four positive clones express the human Fab fragment with antigen specificities.