Studies of the expression, purification, renaturation and biologic activity of an anti-CEA immunotoxin.
- Author:
Hui YANG
1
;
Dan HE
;
Kai CHAO
;
Qing LIN
;
Song YOU
;
Hua-Liang HUANG
Author Information
1. School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110015, China.
- Publication Type:Journal Article
- MeSH:
ADP Ribose Transferases;
biosynthesis;
genetics;
pharmacology;
Antibodies;
genetics;
metabolism;
pharmacology;
Antineoplastic Agents;
metabolism;
pharmacology;
Bacterial Toxins;
biosynthesis;
genetics;
pharmacology;
Carcinoembryonic Antigen;
immunology;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Exotoxins;
biosynthesis;
genetics;
pharmacology;
Humans;
Immunoglobulin Fragments;
biosynthesis;
genetics;
Immunotoxins;
genetics;
isolation & purification;
metabolism;
pharmacology;
Protein Renaturation;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
pharmacology;
Virulence Factors;
biosynthesis;
genetics;
pharmacology
- From:
Chinese Journal of Biotechnology
2004;20(3):348-351
- CountryChina
- Language:Chinese
-
Abstract:
A recombinant immunotoxin named CEA/PE38/KDEL was constructed, which was composed of anti-CEA single-chain Fv and the truncated and modified form of Pseudomonas exotoxin (PE38/KDEL). The CEA/PE38/KDEL immunotoxin was expressed in the E. coli strain BL21 (DE3)-star as inclusion bodies. The denatured inclusion bodies were purified with Ni-NTA chelate agarose, then the constant gradient dialysis was used to perform the refolding of the CEA/PE38/KDEL immunotoxin. Results of FACS and MTT assay indicate that the refolded immunotoxins keep potent and specific cytotoxicity to tumor cells bearing CEA antigens.
