北京大学学报(自然科学版)

从噬菌体抗体库中淘选人绒毛膜促性腺激素的单链抗体

路小龙,张凤英,郭振泉,黄仪秀,朱圣庚   

  1. 北京大学生命科学学院,北京,100871
  • 收稿日期:2001-06-01 出版日期:2002-03-20 发布日期:2002-03-20

Selection of Anti-hCG ScFv from Murine Phage Antibody Library

LU Xiaolong,ZHANG Fengying,GUO Zhenquan,HUANG Yixiu,ZHU Shenggeng   

  1. College of Life Sciences, Peking University, Beijing, 100871
  • Received:2001-06-01 Online:2002-03-20 Published:2002-03-20

摘要: 由鼠源噬菌体抗体库淘选到展示有人绒毛膜促性腺激素单链抗体的噬菌体克隆。用ELISA方法进行检测,从结合力最强的克隆中分离单链抗体基因,插入经改造的高效表达载体pET-21a(+)中,转化大肠杆菌,诱导表达。从培养基中可检测到可溶性抗人绒毛膜促性腺激素的单链抗体。

关键词: 噬菌体抗体库, 人绒毛膜促性腺激素, 单链抗体

Abstract: The murine phage antibody library was amplified and then panned by human chorionic gonadotropin (hCG) for four rounds. The last round enriched phage clones were used to reinfect E coli TG1 and 15 separate colonies were assayed by ELISA. The recombinant phages which were rescued from colony 12# showed the highest affinity to immobilized hCG. The Scfv gene was inserted into the reconstructed pET-21a(+) and expressed in E coli BL21(DE3). The dot blot hybridization verified that the ScFv was secreted to the medium.

Key words: phage antibody library, human chorionic gonadotropin (hCG), single-chain Fv fragment (ScFv)

中图分类号: