Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
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DOI：
10.1097/mbc.0000000000000477
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发表时间：
2016-07-01
影响因子：
1.1
通讯作者：
Thim, Lars
中科院分区：
文献类型：
--
作者：
Ahmadian, Haleh;Hansen, Ernst B.;Thim, Lars
Turoctocog alfa （NovoEight）是在中国仓鼠卵巢细胞中产生的具有截断b结构域的第三代重组因子VIII （rFVIII）。在此过程中不使用人类或动物来源的材料。本研究的目的是描述turoctocog alfa的分子设计和纯化过程。turoctocog alfa采用五步纯化工艺：在混合模式树脂上捕获蛋白质；免疫亲和层析使用独特的，重组生产的抗fviii单抗；阴离子交换色谱法；纳滤和粒径排除色谱。这一过程使诸如宿主细胞蛋白（HCPs）和高分子量蛋白（HMWPs）等杂质减少到非常低的水平。免疫亲和步骤对于去除fviii相关降解产物非常重要。本文所示的生产规模数据证实了纯化工艺的稳健性和杂质的可靠和一致的减少。每个步骤对最终产品纯度的贡献被描述并显示为三个生产批次。Turoctocog alfa是在中国仓鼠卵巢细胞中制造的第三代b结构域截断的rFVIII产品，不使用动物或人类来源的蛋白质。五步纯化过程的结果是均匀的，高纯度的rFVIII产品。版权所有2016威科集团有限公司版权所有。
Turoctocog alfa (NovoEight) is a third-generation recombinant factor VIII (rFVIII) with a truncated B-domain that is manufactured in Chinese hamster ovary cells. No human or animal-derived materials are used in the process. The aim of this study is to describe the molecular design and purification process for turoctocog alfa. A five-step purification process is applied to turoctocog alfa: protein capture on mixed-mode resin; immunoaffinity chromatography using a unique, recombinantly produced anti-FVIII mAb; anion exchange chromatography; nanofiltration and size exclusion chromatography. This process enabled reduction of impurities such as host cell proteins (HCPs) and high molecular weight proteins (HMWPs) to a very low level. The immunoaffinity step is very important for the removal of FVIII-related degradation products. Manufacturing scale data shown in this article confirmed the robustness of the purification process and a reliable and consistent reduction of the impurities. The contribution of each step to the final product purity is described and shown for three manufacturing batches. Turoctocog alfa, a third-generation B-domain truncated rFVIII product is manufactured in Chinese hamster ovary cells without the use of animal or human-derived proteins. The five-step purification process results in a homogenous, highly purified rFVIII product. Copyright (C) 2016 Wolters Kluwer Health, Inc. All rights reserved.