噬菌体单链抗体导向溶栓剂的构建

Fibrin\|specific antibodies were isolated from a phage\|displayed single\|chain antibody(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 S 2444 assay,the uPA activity of the chimera was about 3.1×10 3?IU/mg periplasmic protein of host E.coli .It also kept up the affinity to fibrin.

Construction of Phage Single-chain Antibody-targeted Thrombolytics

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.