[Prokaryotic expression, purification, refolding and biological assays of recombinant human interleukin 4 inclusion body].
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.
Li
Jiong
J
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Cui
Kaijun
K
Wen
Jing
J
Zhao
Zhiwei
Z
Chen
Ping
P
Tian
Ling
L
Kan
Bing
B
Wen
Yanjun
Y
Deng
Hongxin
H
Fan
Linyu
L
Wei
Yuquan
Y
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D015847:Interleukin-4 / Q000096:biosynthesis* / Q000235:genetics; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
C508598:IL4 protein, human; D011994:Recombinant Proteins; D015847:Interleukin-4