In this review, the basic concepts and applications of design of experiments (DoE) in recombinant protein biotechnology will be discussed. The production of recombinant proteins usually begins with the construction of an expression vector that is then introduced into a microbial host. The target protein is overexpressed in the host's cells and subsequently, it is isolated using a suitable purification method, its activity is assessed using a biological assay, while its crystallization is often required. Because each protein is unique and due to the complex interactions among the reagents in experiments, it is impossible that one set of reaction conditions would be optimal for all cases. Optimization of experimental conditions is usually carried out by the inefficient one-factor-at-a-time approach that does not take into account the combined effects of factors on a process. On the other hand, DoE approaches with a carefully selected small set of experiments, and therefore with a reduced cost and in a limited amount of time predict the effect of each factor and the effects of their interactions on a process. Importantly, several software packages are available that facilitate the choice of the DoE approach, design of the experiments, and analysis of the results.

中文翻译：

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实验设计作为重组蛋白生物技术优化的工具：从构建体到晶体。

在这篇综述中，将讨论重组蛋白生物技术中实验设计（DoE）的基本概念和应用。重组蛋白的生产通常从构建表达载体开始，然后将其引入微生物宿主。目标蛋白在宿主细胞中过度表达，随后，使用合适的纯化方法将其分离，并通过生物学分析评估其活性，而经常需要结晶。由于每种蛋白质都是独特的，并且由于实验中试剂之间的复杂相互作用，因此不可能针对所有情况优化一组反应条件。实验条件的优化通常是通过效率低下的一次一因素方法进行的，该方法没有考虑因素对过程的综合影响。另一方面，DoE采用精心挑选的少量实验进行处理，因此以降低的成本并在有限的时间内预测了每个因素的影响及其相互作用对过程的影响。重要的是，有几个软件包可用来简化DoE方法的选择，实验的设计以及结果的分析。