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首页> 外文期刊>Journal of mass spectrometry: JMS >Quick quantification of proteins by MALDI
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Quick quantification of proteins by MALDI

机译：通过MALDI快速定量蛋白质

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Previously, we reported that the matrix-assisted laser desorption ionization spectrum of a peptide became reproducible when an effective temperature was held constant. Using a calibration curve drawn by plotting the peptide-to-matrix ion abundance ratio versus the peptide concentration in a solid sample, a peptide could be quantified without the use of any internal standard. In this work, we quantified proteins by quantifying their tryptic peptides with the aforementioned method. We modified the digestion process; e.g. disulfide bonds were not cleaved, so that hardly any reagent other than trypsin remained after the digestion process. This allowed the preparation of a sample by the direct mixing of a digestion mixture with a matrix solution. We also observed that the efficiency of the matrix-to-peptide proton transfer, as measured by its reaction quotient, was similar for peptides with arginine at the C-terminus. With the reaction quotient averaged over many such peptides, we could rapidly quantify proteins. Most importantly, no peptide standard, not to mention its isotopically labeled analog, was needed in this method. Copyright (c) 2015 John Wiley & Sons, Ltd.
机译：以前，我们报道了当有效温度保持恒定时，肽的基质辅助激光解吸电离光谱变得可重现。使用通过绘制肽与基质离子丰度比对固体样品中肽浓度绘制的校准曲线，可以在不使用任何内标的情况下对肽进行定量。在这项工作中，我们通过使用上述方法对蛋白质的胰蛋白酶解肽进行了定量。我们修改了消化过程;例如二硫键没有被切割，因此在消化过程后几乎没有剩下除胰蛋白酶以外的任何试剂。这允许通过将消化混合物与基质溶液直接混合来制备样品。我们还观察到，通过其反应商测量的基质到肽质子转移的效率与在C端带有精氨酸的肽相似。使用许多此类肽的平均反应商，我们可以快速定量蛋白质。最重要的是，此方法不需要肽标准品，更不用说其同位素标记的类似物。版权所有（c）2015 John Wiley＆Sons，Ltd.

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