Plate-forme de Biophysique Moléculaire, Institut Pasteur, UMR 3528 CNRS, Paris, France.
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Methods Mol Biol. 2021;2263:3-46. doi: 10.1007/978-1-0716-1197-5_1.
One essential prerequisite of any experiment involving a purified protein, such as interaction studies or structural and biophysical characterization, is to work with a "good-quality" sample in order to ensure reproducibility and reliability of the data. Here, we define a "good-quality" sample as a protein preparation that fulfills three criteria: (1) the preparation contains a protein that is pure and soluble and exhibits structural and functional integrity, (2) the protein must be structurally homogeneous, and (3) the preparation must be reproducible. To ensure effective quality control (QC) of all these parameters, we suggest to follow a simple workflow involving the use of gel electrophoresis, light scattering, and spectroscopic experiments. We describe the techniques used in every step of this workflow and provide easy-to-use standard protocols for each step.
任何涉及纯化蛋白的实验的一个基本前提，例如相互作用研究或结构和生物物理特性分析，都是使用“高质量”的样品，以确保数据的可重复性和可靠性。在这里，我们将“高质量”的样品定义为满足三个标准的蛋白质制剂：（1）该制剂包含纯的和可溶的且表现出结构和功能完整性的蛋白质；（2）蛋白质必须具有结构均一性；（3）制剂必须具有重现性。为了确保所有这些参数的有效质量控制 (QC)，我们建议遵循一个简单的工作流程，该流程涉及凝胶电泳、光散射和光谱实验的使用。我们描述了该工作流程每一步中使用的技术，并为每一步提供了易于使用的标准方案。
