Acta Scientiarum Naturalium Universitatis Pekinensis

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Construction of Phage Single-chain Antibody-targeted Thrombolytics

SUN Lei,HUANG Yixiu,ZHU Shenggeng   

  1. College of Life Sciences, Peking University, Beijing, 100871
  • Received:2000-01-07 Online:2000-11-20 Published:2000-11-20

噬菌体单链抗体导向溶栓剂的构建

孙雷,黄仪秀,朱圣庚   

  1. 北京大学生命科学学院,北京,100871

Abstract: Fibrin-specific antibodies were isolated from a phage-displayed single-chainantibody(ScAb) library using affinity selection or panning. DNA shuffling was introduced to realign the specific antibody genes in order to mimic the antibody maturation in vivo. After cloning of shuffling products, a secondary library was constructed, from which the specific antibody against fibrin with higher activity was selected through panning and screening. The fibrin-specific antibody gene and uPA functional domain gene fragment were joined together, then inserted into expression plasmid pET-21a. The ScAb/uPA fusion protein was expressed with the induction of IPTG. In substrate S2444 assay, the uPA activity of the chimera was about 3.1*103 IU/mg periplasmic protein of host E.coli. It also kept up the affinity to fibrin.

Key words: phage display, single-chain antibody, DNA shuffling, targeted thrombolytics, urokinase

关键词: 噬菌体展示:单链抗体, DNA改组, 导向溶栓剂, 尿激酶

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