Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
Institute for Molecular Biotechnology, Worringerweg 1, RWTH Aachen University, Aachen, Germany.
Methods Mol Biol. 2022;2480:295-310. doi: 10.1007/978-1-0716-2241-4_16.
The efficient extraction and purification of recombinant proteins from leaf and seed tissues is often a challenging task, involving multiple steps that must be optimized by identifying and accommodating complex parameter interactions. Conventional one-factor-at-a-time approaches fail to reveal these complex interactions and often result in sub-optimal processes with unnecessary costs. Here, we describe generic considerations to identify global optima for the extraction and purification of recombinant proteins from complex plant matrices. The corresponding experiments can help to streamline downstream processing by reducing the time, costs, and number of unit operations. The procedure involves the knowledge-based selection of factors for screening, the systematic design and analysis of experiments, and the iterative refinement of suitable conditions. The resulting descriptive models can be used to guide process scale-up and offer scientific justifications for process development decisions in negotiations with regulatory authorities.
从叶片和种子组织中高效提取和纯化重组蛋白通常是一项具有挑战性的任务，需要通过识别和适应复杂的参数相互作用来优化多个步骤。传统的单因素逐一方法无法揭示这些复杂的相互作用，并且通常会导致不必要的成本和次优的过程。在这里，我们描述了从复杂植物基质中提取和纯化重组蛋白的全局最优的通用考虑因素。相应的实验可以通过减少时间、成本和单元操作的数量来简化下游处理。该程序涉及基于知识的筛选因素选择、实验的系统设计和分析以及合适条件的迭代改进。所得描述性模型可用于指导工艺放大，并为与监管机构进行的工艺开发决策提供科学依据。
