Effect of Ca2+ and Mg2+ concentration in culture medium on the activation of recombinant factor IX produced in Chinese hamster ovary cells

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Factor IX (FIX) plays an important role in the blood coagulation cascade. When Chinese hamster ovary (CHO) cells producing recombinant human FIX were cultivated using a serum-free medium (SFM) containing 1.12 mM of Ca2+ and 0.82 mM of Mg2+, a significant amount of active FIX (aFIX) was converted into undesirable activated FIX (FIXa) in the later phase of batch culture. In an effort to improve aFIX production from CHO cells, the effect of Ca2+ and Mg2+ concentrations in the culture medium on the activation of aFIX to FIXa was investigated using SFM with various concentrations of Ca2+ and Mg2+ in the range of 0-1.0 mM. The highest aFIX concentration of 1.36 IU/mL. was obtained at 1.0 mM Ca2+ and 1.0mM Mg2+. but the activation of aFIX to FIXa in the later phase of culture was rapid and significant. In contrast at 0.5 mM Ca2+ and 1.0 mM Mg2+, the aFIX concentration of 1.33 IU/mL was obtained and did not decrease significantly in the later phase of culture. Taken together, lowering Ca2+ concentration from 1.0 to 0.5 mM inhibits the activation of aFIX to FIXa in the later phase of culture, fortifying the robustness of downstream bioprocessing. (C) 2009 Elsevier B.V. All rights reserved.
Publisher
ELSEVIER SCIENCE BV
Issue Date
2009-07
Language
English
Article Type
Article
Keywords

GLA DOMAIN; CRYSTAL-STRUCTURE; BINDING-PROTEIN; HEMOPHILIA-B; COAGULATION; EXPRESSION; MEMBRANE; ASSAYS; EDTA; CHO

Citation

JOURNAL OF BIOTECHNOLOGY, v.142, no.3-4, pp.275 - 278

ISSN
0168-1656
DOI
10.1016/j.jbiotec.2009.06.001
URI
http://hdl.handle.net/10203/100921
Appears in Collection
BS-Journal Papers(저널논문)
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