[Purification of recombinant hEGF expressed in yeast Pichia pastoris and the study on its characters].
OBJECTIVE
To obtain recombinant human epidermal growth factor(hEGF) that can be used in animal experiments and clinical trial.


METHODS
Chemically synthesized hEGF gene was expressed in Yeast Pichia pastoris and the secretory hEGF was purified by Phenysepharose 6 Fast Flow(high sub), Q-sepharose High Performance, and Superdex 30 chromatography, and its characters were studied by respective methods.


RESULTS
The purified hEGF doesn't contain pyrogen, endotoxin, or yeast chromosome DNA and the purity reached 98%. The recombinant human EGF has correct molecular weight, pI, N-terminal amino acids sequences, peptide map, ultraviolet spectrum and well-biological activity.


CONCLUSIONS
The purified hEGF is in accord with the requirements for animal experiments and clinical trial which provides the basis of preparing EGF agents for clinical test.
Huang
B R
BR
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China. huangbr@public.bta.net.cn
Cai
L W
LW
Wang
X
X
Ma
X M
XM
Ma
X L
XL
Li
J J
JJ
Li
H Z
HZ
Wei
Z M
ZM
Liang
Q
Q
Zheng
H J
HJ
Tang
Q H
QH
Tang
S X
SX
Liao
H T
HT
Xiang
X Z
XZ
D000818:Animals; D041702:BALB 3T3 Cells; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D051379:Mice; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D004815:Epidermal Growth Factor