DoE based integration approach of upstream and downstream processing regarding HCP and ATPE as harvest operation
DoE based integration approach of upstream and downstream processing regarding HCP and ATPE as harvest operation
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DOI：
10.1016/j.bej.2016.06.016
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发表时间：
2016-09-15
影响因子：
3.9
通讯作者：
Strube, Jochen
中科院分区：
文献类型：
--
作者：
Gronemeyer, Petra;Ditz, Reinhard;Strube, Jochen
传统的 USP 优化概念提高了产品滴度并显着提高了杂质谱。培养液的不同成分给下游加工带来了额外的挑战。这项工作通过结合方法和技术方法来解决由此产生的问题。方法方法基于通过扩展 USP 优化范围来整合上游和下游工艺开发。除了高产品滴度、细胞数量和产品质量外，还考虑了宿主细胞蛋白质的组成和浓度。这项工作证明了这种方法的可行性。使用产生 IgG 的模型 CHO 细胞系来开发培养基，该培养基可产生更高的滴度和细胞数量以及更低的 HCP 量。 HCP 的新组合物旨在简化后续纯化步骤。对实验进行了统计规划并对结果进行了验证，所有这些都导致了更高的产物滴度但更低的 HCP 水平。下一步，研究了培养和细胞收获操作对杂质的影响。评估了离心、过滤和水两相萃取在不损失产物的情况下分离细胞和 HCP 的能力。 ATPE 因其高细胞分离效率、高产量和大量去除 HCP 而成为细胞收获操作的选择，而这是离心和过滤无法实现的。 (C) 2016 Elsevier B.V. 保留所有权利。
Conventional USP optimization concepts have led to increasing product titers and significantly raised impurity profiles. Varying compositions of cultivation broth present additional challenges in downstream processing. This work addresses the resulting problems by combining methodical and technological approaches.The methodical approach is based on the integration of upstream and downstream process development by extending the range of USP optimization. In addition to high product titer, cell numbers and product quality, the composition and concentration of host cell proteins were taken into account. The feasibility of this approach was demonstrated within this work. A model CHO cell line producing IgG was used to develop a medium, which generates higher titers and cell numbers as well as a lower amount of HCP. The new composition of HCP was designed to simplify subsequent purification steps. Experiments were statistically planned and the results were validated, all resulting in higher product titers but lower HCP levels.In the next step, the influence of the cultivation and cell harvest operations on the impurities was investigated. Centrifugation, filtration and aqueous two phase extraction were evaluated regarding their ability to separate cells and HCP without product loss. ATPE distinguished itself as the cell harvest operation of choice by high cell separation efficiency, high yields, and a substantial removal of HCP which could not be met by centrifugation and filtration. (C) 2016 Elsevier B.V. All rights reserved.