Raynal Bertrand, Lenormand Pascal, Baron Bruno, Hoos Sylviane, England Patrick
Institut Pasteur, Biophysics of Macromolecules and their Interactions, 25 rue du Docteur Roux, 75724, Paris Cedex 15, France.
CNRS-UMR3528, Institut Pasteur, Departement of Structural Biology and Chemistry, Paris, France.
Microb Cell Fact. 2014 Dec 30;13:180. doi: 10.1186/s12934-014-0180-6.
Purified protein quality control is the final and critical check-point of any protein production process. Unfortunately, it is too often overlooked and performed hastily, resulting in irreproducible and misleading observations in downstream applications. In this review, we aim at proposing a simple-to-follow workflow based on an ensemble of widely available physico-chemical technologies, to assess sequentially the essential properties of any protein sample: purity and integrity, homogeneity and activity. Approaches are then suggested to optimize the homogeneity, time-stability and storage conditions of purified protein preparations, as well as methods to rapidly evaluate their reproducibility and lot-to-lot consistency.
纯化蛋白质质量控制是任何蛋白质生产过程的最后关键检查点。不幸的是,它常常被忽视且执行仓促,导致下游应用中出现不可重复和误导性的观察结果。在本综述中,我们旨在基于一系列广泛可用的物理化学技术,提出一个易于遵循的工作流程,以依次评估任何蛋白质样品的基本特性:纯度和完整性、均一性和活性。然后提出优化纯化蛋白质制剂均一性、时间稳定性和储存条件的方法,以及快速评估其可重复性和批次间一致性的方法。