目的:探讨重组免疫毒素hdsFv-hEDN在大肠杆菌中的可溶性表达和纯化,观察其体外杀伤作用。方法:利用IPTG诱导hdsFv-hEDN免疫毒素在大肠杆菌中的表达。诱导转化菌经超声碎菌、离心、N i-NTA Agarose亲合层析等步骤纯化,并通过SDS-PAGE和W estern-b lot进行分析及检测。采用MTT比色法观察其对人肝癌细胞的杀伤作用。结果:IPTG诱导后,hdsFv-hEDN在大肠杆菌中获得了可溶性表达,表达量为8%。表达产物纯化后,纯度达到基本均一。MTT结果表明,其对人肝癌细胞具有一定的杀伤作用。结论:本研究实现了hdsFv-hEDN重组免疫毒素在大肠杆菌中的可溶性表达及纯化,获得了具有一定杀伤作用的抗肝癌重组免疫毒素。 OBJECTIVE: To investigate the soluble expression and purification of recombinant immunotoxin hdsFv-hEDN in Escherichia coli and observe its in vitro cytotoxicity. Methods: IPTG was used to induce the expression of hdsFv-hEDN immunotoxin in Escherichia coli. The transformed bacteria were induced to be purified by ultrasonication, centrifugation, N i-NTA Agarose affinity chromatography, and analyzed by SDS-PAGE and Western-blot analysis. MTT colorimetric method was used to observe the killing effect on human hepatoma cells. Results: After induced by IPTG, hdsFv-hEDN was expressed in E. coli with a soluble level of 8%. Purification of the expression product, the purity of the basic uniform. MTT results show that it has a certain killing effect on human hepatoma cells. Conclusion: The htdFv-hEDN recombinant immunotoxin was successfully expressed and purified in Escherichia coli and the anti-HCC recombinant immunotoxin was obtained.