Expression, purification and functional detection of anti-HCC hdsFv fused with RC-RNase

Objective To obtain a new kind of anti-HCC recombinant immunotoxin, which has potentialities for clinical application, high specificity and increased stability. Methods The prokaryotic expression vector pTIH-hdsFv-RC-RNase was induced to express in E.coli by IPTG. The expressed product was purified by Ni-NTA agarose affinity chromatography under native conditions and mildly refolded. The immunocytochemical staining and MTT colorimetry were used respectively to detect the specifically binding activity and ki...