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HERO ID
2283527
Reference Type
Journal Article
Title
Evaluation of butyrate induced production of A Mannose-6-phosphorylated therapeutic enzyme using Parallel Bioreactors
Author(s)
Madhavarao, CN; Agarabi, CD; Wong, L; Müller-Loennies, S; Braulke, T; Khan, M; Anderson, H; Johnson, GR
Year
2014
Is Peer Reviewed?
Yes
Journal
Biotechnology and Applied Biochemistry
ISSN:
0885-4513
EISSN:
1470-8744
Volume
61
Issue
2
Page Numbers
184-192
Language
English
PMID
24033810
DOI
10.1002/bab.1151
Abstract
Bioreactor process changes can have a profound effect on the yield and quality of biotechnology products. Mannose-6-phosphate (M6P) glycan content and the enzymatic catalytic kinetic parameters are critical quality attributes (CQAs) of many therapeutic enzymes used to treat lysosomal storage diseases. Here, we have evaluated the effect of adding butyrate to bioreactor production cultures of human recombinant β-glucuronidase produced from CHO-K1 cells, with an emphasis on CQAs. The β-glucuronidase produced in parallel bioreactors was quantified by capillary electrophoresis, the catalytic kinetic parameters were measured using steady-state analysis, and mannose-6-phosphorylation status was assessed using an M6P-specific single chain antibody fragment. Using this approach we found that butyrate treatment increased β-glucuronidase production up to ∼3-fold without significantly affecting the catalytic properties of the enzyme. However, M6P content in β-glucuronidase was inversely correlated with the increased enzyme production induced by butyrate treatment. This assessment demonstrated that although butyrate dramatically increased β-glucuronidase production in bioreactors, it adversely impacted the mannose-6-phosphorylation of this lysosomal storage disease therapeutic enzyme. This strategy may have utility in evaluating manufacturing process changes to improve therapeutic enzyme yields and CQAs.
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n-Butanol
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Pubmed
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Pubmed - 3/2014
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Use in sample prep or assay
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