Simple fabrication of hydrophobic surface target for increased sensitivity and homogeneity in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of peptides, phosphopeptides, carbohydrates and proteins
Simple fabrication of hydrophobic surface target for increased sensitivity and homogeneity in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of peptides, phosphopeptides, carbohydrates and proteins
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DOI：
10.1016/j.aca.2013.04.029
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发表时间：
2013-06-14
影响因子：
6.2
通讯作者：
Tsai, Fuu-Jen
中科院分区：
文献类型：
--
作者：
Chen, Chao-Jung;Lai, Chien-Chen;Tsai, Fuu-Jen
为了在基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF)分析中增强样品信号，改善样品的均匀性，本研究建立了一种简单、快速、高效的样品制备方法。聚二甲基硅氧烷(PDMS)涂覆在不锈钢MALDI板上，形成透明、疏水的表面，增强了样品信号，而不产生明显的背景信号。与使用未经修饰的商用金属MALDI板相比，由于PDMS涂层板的样品斑点面积减少，肽信号增强了7.1-11.0倍，并显示了改进的肽质量指纹图谱(PMF)和MS/MS肽测序结果。在以2，5-二羟基苯甲酸(DHB)为基质的磷酸和碳水化合物的分析中，涂覆PDMS的平板样品均匀度和信号增强分别提高了5.2-8.2倍和2.8-3.2倍。通过MS/MS分析，提高了观察更多独特碎片离子的灵敏度，从而成功地区分了同分异构体碳水化合物。在用芥子酸(SA)基质进行蛋白质分析时，观察到信号增强了3.4倍。PDMS薄膜涂层易于去除和重新填充，避免了样品携带，适用于各种商业MALDI板。PDMS涂层方法是一种简单、实用和有吸引力的增强分析物信号和均一性的方法。(C)2013爱思唯尔B.V.保留所有权利。
To enhance sample signals and improve homogeneity in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis, a simple, rapid, and efficient sample preparation method was developed in this study. Polydimethylsiloxane (PDMS) was coated on a stainless steel MALDI plate, forming a transparent, hydrophobic surface that enhanced sample signals without producing observable background signals. Compared to the use of an unmodified commercial metal MALDI plate, peptide signals were enhanced by similar to 7.1-11.0-fold due to the reduced sample spot area of the PDMS-coated plate, and showed improved peptide mass fingerprinting (PMF) and MS/MS peptide sequencing results. In the analysis of phosphopeptides and carbohydrates with a 2,5-dihydroxybenzoic acid (DHB) matrix, the PDMS-coated plate showed improved sample homogeneity and signal enhancements of similar to 5.2-8.2-fold and similar to 2.8-3.2-fold, respectively. Improved sensitivity in the observation of more unique fragment ions by MS/MS analysis, to successfully distinguish isomeric carbohydrates, was also illustrated. In protein analysis with a sinapinic acid (SA) matrix, a 3.4-fold signal enhancement was observed. The PDMS film coating was easily removed and refabricated to avoid sample carryover, and was applicable to diverse commercial MALDI plates. The PDMS-coated approach is a simple, practical, and attractive method for enhancing analyte signals and homogeneity. (C) 2013 Elsevier B.V. All rights reserved.