Effect of Ca2+ and Mg2+ concentration in culture medium on the activation of recombinant factor IX produced in Chinese hamster ovary cells.
Effect of Ca2+ and Mg2+ concentration in culture medium on the activation of recombinant factor IX produced in Chinese hamster ovary cells.
复制标题
培养基中 Ca2 和 Mg2 浓度对中国仓鼠卵巢细胞产生的重组因子 IX 活化的影响。
DOI：
10.1016/j.jbiotec.2009.06.001
复制
发表时间：
2009
影响因子：
4.1
通讯作者：
G. Lee
中科院分区：
文献类型：
--
作者：
Won Hee Kim;Jung;Yeup Yoon;G. Lee
因子 IX (FIX) 在凝血级联中发挥着重要作用。当使用含有1.12mM Ca2+和0.82mM Mg2+的无血清培养基（SFM）培养产生重组人FIX的中国仓鼠卵巢（CHO）细胞时，在分批培养的后期阶段，大量活性FIX（aFIX）转化为不需要的活化FIX（FIXa）。为了提高 CHO 细胞的 aFIX 产量，使用具有 0-1.0 mM 范围内不同浓度的 Ca2+ 和 Mg2+ 的 SFM 研究了培养基中 Ca2+ 和 Mg2+ 浓度对 aFIX 活化为 FIXa 的影响。在1.0mM Ca2+和1.0mM Mg2+时获得最高aFIX浓度1.36IU/mL，但在培养后期aFIX向FIXa的激活是快速且显着的。相比之下，在0.5mM Ca2+和1.0mM Mg2+下，aFIX浓度为1.33IU/mL，并且在培养后期没有明显下降。总而言之，将 Ca2+ 浓度从 1.0 降低至 0.5 mM 可抑制培养后期 aFIX 活化为 FIXa，从而增强下游生物加工的稳健性。
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 Ca2+and 0.82mM 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 0–1.0mM. The highest aFIX concentration of 1.36IU/mL was obtained at 1.0mM 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 0.5mM Ca2+and 1.0mM Mg2+, the aFIX concentration of 1.33IU/mL was obtained and did not decrease significantly in the later phase of culture. Taken together, lowering Ca2+concentration from 1.0 to 0.5mM inhibits the activation of aFIX to FIXa in the later phase of culture, fortifying the robustness of downstream bioprocessing.
中国仓鼠卵巢细胞中合成的重组 γ-羧化因子 IX 的表达、纯化和表征。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kaufman,RJ;Wasley,LC;Furie,BC;Furie,B;Shoemaker,CB
通讯作者：
Shoemaker,CB