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首页> 美国卫生研究院文献>Membranes >Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium
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Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium

机译：溶瘤性麻疹病毒浓度的切向流过滤：过滤器性能和细胞培养基的影响

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摘要

The therapeutic use of oncolytic measles virus (MV) for cancer treatment requires >10 infectious MV particles per dose in a highly pure form. The concentration/purification of viruses is typically achieved by tangential flow filtration (TFF) but the efficiency of this process for the preparation of MV has not been tested in detail. We therefore investigated the influence of membrane material, feed composition, and pore size or molecular weight cut-off (MWCO) on the recovery of MV by TFF in concentration mode. We achieved the recovery of infectious MV particles using membranes with a MWCO ≤ 300 kDa regardless of the membrane material and whether or not serum was present in the feed. However, serum proteins in the medium affected membrane flux and promoted fouling. The severity of fouling was dependent on the membrane material, with the cellulose-based membrane showing the lowest susceptibility. We found that impurities such as proteins and host cell DNA were best depleted using membranes with a MWCO ≥ 300 kDa. We conclude that TFF in concentration mode is a robust unit operation to concentrate infectious MV particles while depleting impurities such as non-infectious MV particles, proteins, and host cell DNA.
机译：溶瘤性麻疹病毒（MV）在癌症治疗中的治疗用途需要每剂高纯度形式的> 10个感染性MV颗粒。病毒的浓缩/纯化通常通过切向流过滤（TFF）实现，但尚未详细测试该方法制备MV的效率。因此，我们研究了膜材料，进料组成，孔径或截留分子量（MWCO）对浓缩模式下TFF回收MV的影响。我们使用MWCO≤300 kDa的膜实现了传染性MV颗粒的回收，无论膜的材料如何，饲料中是否存在血清。但是，培养基中的血清蛋白会影响膜通量并促进结垢。结垢的严重程度取决于膜材料，纤维素基膜显示出最低的敏感性。我们发现，使用MWCO≥300 kDa的膜可以最好地消除蛋白质和宿主细胞DNA等杂质。我们得出的结论是，浓缩模式下的TFF是浓缩感染性MV颗粒同时消耗诸如非感染性MV颗粒，蛋白质和宿主细胞DNA等杂质的强大单元操作。

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