Department of Biotechnology and Food Engingeering, Tecnológico de Monterrey, Monterrey, Nuevo León, Mexico.
PLoS One. 2012;7(2):e31438. doi: 10.1371/journal.pone.0031438. Epub 2012 Feb 17.
The identification of proteins by mass spectrometry is a standard method in biopharmaceutical quality control and biochemical research. Prior to identification by mass spectrometry, proteins are usually pre-separated by electrophoresis. However, current protein staining and de-staining protocols are tedious and time consuming, and therefore prolong the sample preparation time for mass spectrometry.
We developed a 1-minute covalent pre-gel staining protocol for proteins, which does not require de-staining before the mass spectrometry analysis. We investigated the electrophoretic properties of derivatized proteins and peptides and studied their behavior in mass spectrometry. Further, we elucidated the preferred reaction of proteins with Uniblue A and demonstrate the integration of the peptide derivatization into typical informatics tools.
The Uniblue A staining method drastically speeds up the sample preparation for the mass spectrometry based identification of proteins. The application of this chemo-proteomic strategy will be advantageous for routine quality control of proteins and for time-critical tasks in protein analysis.
通过质谱法鉴定蛋白质是生物制药质量控制和生化研究中的标准方法。在通过质谱法进行鉴定之前，通常通过电泳对蛋白质进行预分离。然而，目前的蛋白质染色和脱色方案繁琐且耗时，因此延长了用于质谱分析的样品制备时间。
我们开发了一种 1 分钟的共价预胶染色方案，用于蛋白质，在进行质谱分析之前不需要进行脱色。我们研究了衍生化蛋白质和肽的电泳性质，并研究了它们在质谱中的行为。此外，我们阐明了蛋白质与 Uniblue A 的优选反应，并证明了肽衍生化与典型信息学工具的集成。
Uniblue A 染色方法极大地加快了基于质谱法鉴定蛋白质的样品制备速度。这种化学蛋白质组学策略的应用将有利于蛋白质的常规质量控制和蛋白质分析中的时间关键任务。
