A three-step purification method of large quantities of human recombinant α endothelial cellular growth factor for clinical use

• Published in: International journal of molecular medicine 2007-01, Vol.19 (1), p.97-103
• Main Authors: Sauter, Alexander, Lambert, Katherine, Rupf, Ann-Katrin, Von Specht, Bernd-Ulrich, Hörmann, Karl, Naim, Ramin
• Format: Article
• Language: English
• Subjects: Animals; Chromatography, Affinity - methods; Chromatography, Ion Exchange - methods; Cricetinae; DNA - biosynthesis; Endothelial Growth Factors - isolation & purification; Endothelial Growth Factors - pharmacology; Humans; In Vitro Techniques; Membranes - drug effects; Ovum - chemistry; Polymyxin B - chemistry; Sepharose - analogs & derivatives; Sepharose - chemistry; Transformation, Bacterial
• ISSN: 1107-3756; 1791-244X
• DOI: 10.3892/ijmm.19.1.97

The endothelial cellular growth factor α-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 α-ECGF was expressed in E. coli. Isolation of rh α-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 α-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.