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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/11175

Title: Enhancing Production of Recombinant BMP-2 in Mammalian Cell Culture Systems by Inhibition of Pro-protein Cleavage using 9DR Peptides
Authors: Zhou, Jing-Jing Aileen
Advisor: Cameron, Clokie
Sean, Peel
Department: Dentistry
Keywords: recombinant BMP-2 production
furin cleavage
Issue Date: 30-Jul-2008
Abstract: Introduction: Mammalian cell recombinant bone morphogenetic protein (rBMP) synthesis is reported to be poor. The BMP pro-domain may be involved in folding, stability and secretion. Objectives: Investigate the effect of inhibition of pro-domain cleavage on rhBMP-2 production. Methods: CHO cells transfected with human BMP-2 (hBMP-2) were cultured in the presence of the proprotein convertase inhibitor 9DR in short (multi-well) and long-term (bioreactor) cultures. Mature and proBMP secretion was measured by ELISA and characterized by Western blot. BMP activity was determined by C2C12 bioassay. Results: 9DR significantly enhanced the yields of both pro- and mature hBMP-2 in short and long-term cultures, without any negative effects on cell growth or viability. The rhBMP-2 was biologically active. ProBMP-2 could be converted by exogenous furin treatment into mature BMP-2 as shown by Western blot. Conclusions: 9DR increases rhBMP-2 production and is a simple, but effective way to enhance the yield of active, mature BMP-2 in CHO cells.
URI: http://hdl.handle.net/1807/11175
Appears in Collections:Master
Faculty of Dentistry - Master theses

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