Prokaryotic expression, purification, refolding and biological assays of recombinant human interleukin 4 inclusion body.
- Author:
Jiong LI
1
;
Kaijun CUI
;
Jing WEN
;
Zhiwei ZHAO
;
Ping CHEN
;
Ling TIAN
;
Bing KAN
;
Yanjun WEN
;
Hongxin DENG
;
Linyu FAN
;
Yuquan WEI
Author Information
1. State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
Humans;
Inclusion Bodies;
metabolism;
Interleukin-4;
biosynthesis;
genetics;
Protein Folding;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification
- From:
Journal of Biomedical Engineering
2007;24(4):866-869
- CountryChina
- Language:Chinese
-
Abstract:
A DNA fragment encoding human interleukin 4 was obtained by PCR from pORF-hIL4 plasmid. The amplified fragment was inserted into prokaryotic expression vector PQE60 and recombinant protein was expressed in E. Coli M15 by adding isopropyl-beta-D-thiogalactoside (IPTG). The hIL-4 protein was present as insoluble inclusion bodies in the bacterial extract. After denaturation of inclusion bodies with 5 mol/L guanidine hydrochloride, the supernate was diluted to get renaturized. Then dialysis and Ni chelating chromatography were used for purification. TF-1 proliferation assay of recombinant human interleukin 4 was performed, and then rhIL-4 was fit to be used for proliferation of human dendritic cells from monocyte in vitro.
