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Biotechnol Appl Biochem 2014 Mar;61(2):184-92

Evaluation of butyrate-induced production of a mannose-6-phosphorylated therapeutic enzyme using parallel bioreactors.

Madhavarao CN, Agarabi CD, Wong L, Muller-Loennies S, Braulke T, Khan M, Anderson H, Johnson GR


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 beta-glucuronidase produced from CHO-K1 cells, with an emphasis on CQAs. The beta-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 beta-glucuronidase production up to approximately 3-fold without significantly affecting the catalytic properties of the enzyme. However, M6P content in beta-glucuronidase was inversely correlated with the increased enzyme production induced by butyrate treatment. This assessment demonstrated that although butyrate dramatically increased beta-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.

Category: Journal Article
PubMed ID: #24033810 DOI: 10.1002/bab.1151
Includes FDA Authors from Scientific Area(s): Drugs
Entry Created: 2013-09-17 Entry Last Modified: 2014-06-22