Meitz Andrea, Sagmeister Patrick, Langemann Timo, Herwig Christoph
Research Center of Pharmaceutical Engineering GmbH, Inffeldgasse 13, 8010 Graz, Austria.
Institute of Biochemical Engineering, Vienna University of Technology, Gumpendorferstrasse 1A/166-4, 1060 Vienna, Austria.
Bioengineering (Basel). 2014 Oct 24;1(4):213-230. doi: 10.3390/bioengineering1040213.
The development, optimization, and analysis of downstream processes are challenged by a high number of potentially critical process parameters that need to be investigated using lab-scale experiments. These process parameters are spread across multiple unit operations and potentially show interactions across unit operations. In this contribution, we present a novel strategy for bioprocess development that considers the risk of parameter interactions across unit operations for efficient experimental design. A novel risk assessment tool (interaction matrix) is introduced to the Quality by Design (QbD) workflow. Using this tool, the risk of interaction across unit operations is rated. Subsequently, a design of experiments (DoE) across unit operations is conducted that has the power to reveal multivariate interdependencies. The power of the presented strategy is demonstrated for protein isolation steps of an inclusion body process, focusing on the quality attribute inclusion body purity. The concentration of Triton X-100 in the course of inclusion body (IB) purification was shown to interact with the g-number of the subsequent centrifugation step. The presented strategy targets a holistic view on the process and allows handling of a high number of experimental parameters across unit operations using minimal experimental effort. It is generically applicable for process development along QbD principles.
下游工艺的开发、优化和分析面临着大量潜在的关键工艺参数的挑战，这些参数需要通过实验室规模的实验进行研究。这些工艺参数分布在多个单元操作中，并且可能在单元操作之间呈现相互作用。在本论文中，我们提出了一种新的生物工艺开发策略，该策略在高效实验设计中考虑了单元操作之间参数相互作用的风险。一种新型风险评估工具（相互作用矩阵）被引入到质量源于设计（QbD）工作流程中。使用该工具，对单元操作之间的相互作用风险进行评级。随后，进行跨单元操作的实验设计（DoE），其能够揭示多变量之间的相互依存关系。以包涵体工艺的蛋白质分离步骤为例，重点关注质量属性包涵体纯度，展示了所提出策略的有效性。结果表明，在包涵体（IB）纯化过程中Triton X-100的浓度与后续离心步骤的g值存在相互作用。所提出的策略旨在对工艺进行整体考量，并允许以最少的实验工作量处理跨单元操作的大量实验参数。它普遍适用于遵循QbD原则的工艺开发。
