Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstraße 1, 39106 Magdeburg, Germany.
J Chromatogr A. 2013 Sep 13;1307:99-110. doi: 10.1016/j.chroma.2013.07.081. Epub 2013 Jul 26.
Continuous size exclusion chromatography for the separation of cell culture-derived influenza virus from contaminating proteins was established successfully. Therefore, an open loop simulated moving bed (SMB) setup with one column per zone was applied. Several operating conditions were tested and overall trends were found to be in agreement with expectations derived from theory. Furthermore, the separation performance was compared to an optimized conventional batch chromatography. The yield of influenza virus in the product fraction, based on a hemagglutination assay, was 70% (SMB) and 80% (batch), respectively. The amount of contaminating protein per product was 0.61μgkHAU(-1) (SMB) compared to 0.29μgkHAU(-1) (batch). This corresponds to a reduction of the respective amount in the feed solution by 60% and 80%, respectively. For both processes, the estimated amount of total protein per vaccine dose would meet the level required for manufacturing of human influenza vaccines prepared in cell cultures. Depending on the strategy chosen for sanitization and equilibration of columns the calculated overall productivity for the SMB process was up to 3.8 times higher compared to the batch mode. SMB, therefore, has the potential to replace single column discontinuous chromatography in order to design more efficient purification trains for production of cell culture-derived influenza vaccines.
成功建立了用于从污染蛋白中分离细胞培养衍生流感病毒的连续排阻色谱法。因此，应用了每个区一个柱的开环模拟移动床（SMB）设备。测试了几种操作条件，发现总体趋势与理论预期相符。此外，还比较了分离性能与优化的常规批处理色谱法。基于血凝（HA）测定的产物部分中流感病毒的产率分别为 70%（SMB）和 80%（批处理）。产物中每单位血凝单位（kHAU）的污染蛋白量分别为 0.61μg（SMB）和 0.29μg（批处理）。这相当于进料溶液中相应物质的量分别减少了 60%和 80%。对于这两种工艺，估计每剂疫苗的总蛋白量都将达到细胞培养制备人用流感疫苗的要求。根据选择的柱消毒和平衡策略，SMB 工艺的计算整体生产率比批处理模式高 3.8 倍。因此，SMB 有可能替代单柱不连续色谱法，以便为细胞培养衍生流感疫苗的生产设计更有效的纯化工艺。
