Expression of biologically active,mature human granulocyte-macrophage colony stimulating factor with anE. coli secretory expression system |
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| Authors: | Robert Greenberg Daniel Lundell Yair Alroy Susan Bonitz Russell Condon James Fossetta Beth Frommer Keith Gewain Mitchell Katz Paul J Leibowitz Dr satwant K Narula Robert Kastelein Anita Van Kimmenade |
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| Institution: | (1) Schering Corporation, 60 Orange Street, 07003 Bloomfield, New Jersey, USA;(2) DNAX Research Institute of Molecular and Cellular Biology, Inc., Palo Alto, California, USA;(3) Present address: Merck, Sharpe & Dohme Research Laboratories, P. O. Box 2000, 07065-0900 Rahway, NJ, USA |
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| Abstract: | Human granulocyte-macrophage colony stimulating factor (HuGM-CSF) was expressed periplasmically inEscherichia coli with the secretory vector, pINIIIompA2 10]. HuGM-CSF protein thus expressed was shown to be faithfully cleaved and biologically active. This protein, however, could not be released by osmotic shock and, on subcellular fractionation, co-sedimented with the outer membrane fraction. The effect of promoters, vectors, host strains, induction conditions and media formulation on expression levels was also evaluated. Some of these factors play a significant role in determining maximal achievable levels of HuGM-CSF in the secretory expression system ofE. coli.Both may be considered first authors. |
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