| abstract |
The invention relates to a rapid purification method for gastric cancer-targeted recombinant immunotoxin, which belongs to the field of bioengineering. Using the constructed and screened recombinant Escherichia coli strain that can highly express soluble rG17PE38KDEL, optimize the large-volume culture conditions, expand the culture induction volume to 1L, inoculate 1% inoculum in LB medium, and add 0.5mM induction after shaking at 37°C for 2 hours IPTG, 180rpm/min 16°C shaking culture for 16h, more than 65% of the expressed protein was in soluble state; after the soluble protein was obtained by ultrasonication, the recombinant toxin rG17PE38KDEL with higher purity and activity was obtained by three-step chromatography, and obtained A high-purity toxin with good activity that can be used for drug evaluation. The purity of the gastric cancer-targeting recombinant immunotoxin rG17PE38KDEL obtained after the purification process can reach more than 95%. After purification, the recombinant toxin rG17PE38KDEL can bind to CCK2R on the surface of tumor cells and kill this kind Target tumor cells. |