通过变性超滤和液相色谱-基质辅助激光解吸电离飞行时间质谱法分析肽段
Analysis of peptides by denaturing ultrafiltration and LC-MALDI-TOF-MS.
作者信息
An Yanming, Goldman Radoslav
机构信息
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA.
出版信息
Methods Mol Biol. 2013;1023:13-9. doi: 10.1007/978-1-4614-7209-4_2.
Abstract
The dynamic range of complex biological samples represents a challenge for mass spectrometric -characterization. Removal of high abundant proteins is a prerequisite for a successful mass spectrometric analysis of low abundant analytes. In particular, plasma and serum proteome span at least ten orders of magnitude and represent a major challenge for biomarker discovery. Immunoaffinity depletion is the most common methods of removal of high abundant proteins. Here we describe coupling of denaturing ultrafiltration, an alternative depletion strategy, with reverse-phase fractionation and mass spectrometry for characterization of low-molecular-weight proteins and peptides.
摘要
复杂生物样品的动态范围对质谱表征而言是一项挑战。去除高丰度蛋白质是成功对低丰度分析物进行质谱分析的先决条件。尤其是血浆和血清蛋白质组跨越至少十个数量级，这对生物标志物的发现构成了重大挑战。免疫亲和去除是去除高丰度蛋白质最常用的方法。在此，我们描述了将变性超滤（一种替代去除策略）与反相分级分离和质谱联用，用于表征低分子量蛋白质和肽。
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