摘要
目的制备对人肝癌细胞具有特异性杀伤活性的抗肝癌重组免疫毒素。方法利用大肠杆菌系统表达抗肝癌hdsFvhEDN基因重组免疫毒素。表达产物经NiNTA亲和层析纯化后,进行特异性杀伤活性检测。结果抗肝癌hdsFvhEDN重组免疫毒素以可溶性形式表达于大肠杆菌培养上清中,并获得有效纯化。ELISA法检测证实其具有与相应抗原特异性结合的活性。细胞毒试验表明对肝癌细胞具有杀伤作用,而对正常肝细胞无损伤。结论已成功地制备了具有特异性结合和杀伤活性的抗肝癌hdsFvhEDN基因重组免疫毒素,为其进一步应用奠定基础。
Objective To prepare a novel recombinant immunotoxin with specific killing activity to human hepatocarcinoma.Methods Recombinant immunotoxin hdsFv-hEDN against hepatocarcinoma was expressed by using E.coli expression system,and the expressed product was purified by Ni-NTA affinity chromatography and determined for specific killing activity.Results Recombinant immunotoxin hdsFv-hEDN against hepatocarcinoma was expressed in a form of soluble protein and purified effectively.ELISA proved the specific binding capacity of expressed product to the corresponding antigen.Cytotoxicity test showed the killing activity of expressed product to hepatocarcinoma cells but no damage to normal hepatic cells.Conclusion The recombinant immunotoxin hdsFv-hEDN with specific binding capacity and killing activity to hepatocarcinoma cells was successfully prepared.It laid a foundation of further application of immunotoxin hdsFv-hEDN.
出处
《中国生物制品学杂志》
CAS
CSCD
2006年第3期220-222,共3页
Chinese Journal of Biologicals
基金
国家自然科学基金资助(30370810).
