Reconstruction of anti-hepatocellular carcinoma scFv pharmacy and its killing effect on hepatocardnoma cell in vitro
- VernacularTitle:抗肝癌单链抗体药剂学改造及其对人类肝癌细胞的体外杀伤实验
- Author:
Guihong ZHANG
;
Yanfang LIU
;
Yong FU
;
Haiyang HU
;
Dawei CHEN
- Publication Type:Journal Article
- Keywords:
Immunotoxin;
Immunoliposome;
Liposome;
Colorimetry
- From:
Cancer Research and Clinic
2008;20(12):802-804
- CountryChina
- Language:Chinese
-
Abstract:
objective To observe whether the killing effect on HCC SMMC-7721 cell of the antihepatocellular carcinoma scFv reconstructed by pharmacy was enhanced or not.Methods Prokarycytic expression vector containing PET32a-RC-RNase was induced to express by IPTG.The inclusion body purified and Western-blotting was used.PC.CHOL and CHS was added in chloroform.Dry membrane was formed after chloroform was removed.RC-RNase protein solution was added to dissolute the membrane.Then pass the solution over a Sephadex G-50 column after ultrasound and filtrated to detect the encapsulation efficiency of the liposome.The solution reacted in EDC.SSNHS and MES for 30 minutes.Then add hdscFv to the solution in 4 ℃ over night.MTT method was used to detect the killing effect on HCC cell of immunoliposome RC-RNase,immunotoxin RC-RNase and liposome RC-RNase in vitro.Resuits The killing effect on HCC cell of immunoliposome RC-RNase is the best.but that of Iiposome RC-RNase is the worst.The respective JC50 are:3.28μg/ml,22.44μg/ml and 98.26μg/ml.Conclusion The anti-hepatocellular carcinoma scFv relomtructed by pharmacy can promote the killing effect on HCC cell and may have potential in the treatment of hepatocarcinoma.
