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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 1.12mM of Ca 2+ and 0.82mM of Mg 2+ , a significant amount of active FIX (aFIX) was converted into undesirable activated FIX (FIXa) in the later phase of batch culture. In an effort...