摘要
以Ｅ．ｃｏｌｉＪ５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．ｃｏｌｉＪ５株结合的阳性克隆，且阳性克隆结合抗原的活性可分．别被ＥｃｏｌｉＪ５株、Ｅ．ｃｏｌｉＲｃ－ＬＰＳ和抗Ｅ．ｃｏｌｉＪ５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．
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.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1996年第3期245-250,共6页
Progress In Biochemistry and Biophysics
关键词
噬菌体抗体库
人源抗体
制备
PCR
phage antibody library
human antibody
