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Updated: Jul 18, 2026
Isolation and Large Scale Expansion of Adult Human Endothelial Colony Forming Progenitor Cells
Published on: October 28, 2009
A three-step purification method of large quantities of human recombinant alpha endothelial cellular growth factor
Alexander Sauter1, Katherine L Lambert, Ann-Katrin Rupf
1Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Mannheim, D-68135 Mannheim, Germany. alexander.sauter@hno.ma.uni-heidelberg.de
Abstract:
The endothelial cellular growth factor alpha-ECGF is a candidate drug for the induction of therapeutic neoangiogenesis. Its use in extensive experimental and clinical trials is hampered by the fact that currently published purification procedures allow only small yields, and the absence of pyrogenic impurities is not demonstrated. The rh alpha-ECGF was expressed in E. coli. Isolation of rh alpha-ECGF from E. coli lysates to apparent homogenicity was achieved by a three step purification procedure involving ionic exchange, heparin-sepharose and polymyxin B chromatography. By this method, 200 mg of rh alpha-ECGF was purified from 15 g wet weight E. coli bacteria. The isolated protein of 18 kDa appeared as a single band after SDS gel electrophoresis and subsequent silver-staining. The biological activity was expressed in the chorion-allantois-membrane assay and in the 3H-thymidine proliferation in baby hamster kidney cells. Drug trials with rabbits revealed no increase in body temperature after intravenous injections with 1 mg rh-ECGF.
