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Overexpression of PACEsol improves BMP-7 processing in recombinant CHO cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
23319 43431 2013 4 PDF Available
Title
Overexpression of PACEsol improves BMP-7 processing in recombinant CHO cells
Abstract

Bone morphogenetic protein-7 (BMP-7) is a member of the TGF-β superfamily and plays a critical role in cartilage, bone, and kidney development. BMP-7 is synthesized as a large precursor and undergoes proteolytic cleavage by subtilisin-like proprotein convertase to secrete the functionally active mature dimer. When CHO cells producing recombinant human BMP-7 (rhBMP) (CHO-BMP-7) were cultivated in a serum-free suspension culture, a significant amount of unwanted precursor forms of rhBMP-7 (ca. 69% of total rhBMP-7), along with the mature form of rhBMP-7, was secreted into the culture medium, likely due to the insufficient amount of the proteolytic cleaving enzyme within the secretory pathway. In order to solve this problem, a soluble form of the paired basic amino acid cleaving enzyme (PACEsol), responsible for the majority of the processing events occurring in the constitutive secretory pathway in mammalian cells, was overexpressed in CHO-BMP-7 cells. Overexpression of PACEsol was effective in processing the precursor forms of BMP-7, while it did not significantly affect cell growth. As a result, the culture supernatants of CHO-BMP-7 cells overexpressing PACEsol contained almost 100% of the mature BMP-7 form. Taken together, the results show that PACEsol overexpression in CHO-BMP-7 cells is an efficient means of increasing the production of mature BMP-7 and facilitating the downstream purification steps by eliminating the need to remove the precursor forms.

► CHO cells producing recombinant BMP-7 were cultivated in a suspension culture. ► A significant amount of unwanted precursor rBMP-7 was secreted into the medium. ► BMP-7 undergoes proteolytic cleavage by PACEsol to become the mature dimer. ► Overexpression of PACEsol was effective in processing the precursor forms of BMP-7.

Keywords
BMP-7; BMP-7 precursor; CHO cells; PACEsol
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Overexpression of PACEsol improves BMP-7 processing in recombinant CHO cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 164, Issue 2, March 2013, Pages 336–339
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us