Production and purification of recombinant human granulocyte–macrophage colony stimulating factor (GM-CSF) from high cell density cultures of Pichia pastoris.

Autor:	Palash Bhatacharya, Gaurav Pandey, Krishna Mukherjee
Zdroj:	Bioprocess & Biosystems Engineering; Sep2007, Vol. 30 Issue 5, p305-312, 8p
Abstrakt:	Abstract  Granulocyte–macrophage colony stimulating factor (GM-CSF) is a hematopoietic growth factor, which has been used as a therapeutic agent in clinical cases like neutropenia. In this study, we report the production of recombinant human GM-CSF in the methylotrophic yeast Pichia pastoris through secretory expression using the inducible AOX1 promoter. Recombinant P. pastoris GS115 cells were grown in fed batch cultures to obtain a biomass density of 55.6 gDCW L−1 and a high volumetric activity of 131 mg L−1 of GM-CSF. The protein migrated as a diffuse band on SDS-PAGE at the range of 28–35 kDa indicating differential glycosylation. The secreted protein was purified to 95% in two steps using cation exchange and size exclusion chromatography. [ABSTRACT FROM AUTHOR]
Databáze:	Complementary Index
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