Solid-phase extraction and elution on diamond (SPEED): A fast and general platform for proteome analysis with mass spectrometry
Solid-phase extraction and elution on diamond (SPEED): A fast and general platform for proteome analysis with mass spectrometry
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DOI：
10.1021/ac052085y
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发表时间：
2006-06-15
影响因子：
7.4
通讯作者：
Chang, Huan-Cheng
中科院分区：
文献类型：
--
作者：
Chen, Wei-Hao;Lee, Sheng-Chung;Chang, Huan-Cheng
提出了一种基于表面功能化金刚石纳米晶的固相萃取洗脱平台。与传统方法相比，该平台有助于纯化和浓缩完整蛋白质及其酶消化物，以便在无需事先去除吸附剂的情况下进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳或基质辅助激光解吸/电离质谱分析。涉及二硫键还原、游离半胱氨酸残基保护、洗去残留化学品和吸附蛋白质的蛋白水解消化的一锅法工作流程可直接在颗粒上进行。该蛋白质后处理平台的实用性和多功能性已被证明与液相色谱-电喷雾串联质谱在人尿液蛋白质组分析。每个尿样的蛋白质组分析可在8 h内完成。
This paper presents a new solid-phase extraction and elution platform based on surface-functionalized diamond nanocrystallites. Compared with conventional methods, the platform facilitates purification and concentration of intact proteins and their enzymatic digests for ensuing sodium dodecyl sulfate-polyacrylamide gel electrophoresis or matrix-assisted laser desorption/ionization mass spectrometry analysis without prior removal of the adsorbent. One-pot work flow involving reduction of disulfide bonds, protection of free cysteine residues, washing off residual chemicals, and proteolytic digestion of adsorbed proteins can be performed directly on the particles. The utility and versatility of this protein workup platform were demonstrated with liquid chromatography-electrospray ionization tandem mass spectrometry in proteome analysis of human urine. The proteome analysis of each urine sample can be completed in 8 h.