Three-layer matrix/sample preparation method for MALDI MS analysis of low nanomolar protein samples
Three-layer matrix/sample preparation method for MALDI MS analysis of low nanomolar protein samples
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DOI：
10.1016/j.jasms.2006.02.012
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发表时间：
2006-06-01
影响因子：
3.2
通讯作者：
Li, Liang
中科院分区：
文献类型：
--
作者：
Keller, Bernd O.;Li, Liang
提出了一种稳定而灵敏的样品制备方法，用于基质辅助激光解吸电离(MALDI)对含有大量常见盐和缓冲液的低纳摩尔浓度蛋白质的分析。这种方法包括通过在MALDI靶上制备的基质种子层上沉积基质溶液来生产密集堆积的亚微米基质晶体。然后将亚微升等量的分析物溶液直接添加到基质晶体的顶部，形成薄层。使用α-氰基-4-羟基肉桂酸(4-HCCA)作为基质，其性能优于其他常用基质，如2，5-二羟基苯甲酸(DHB)、2-(4-羟基-苯偶氮)苯甲酸(HABA)或芥子酸。这种三层法被证明优于其他MALDI样品制备方法，特别是在处理含有盐和缓冲液的低纳摩尔蛋白质溶液时。(J am Soc MS Spectrm 2006，17,780-785)(C)2006美国质谱学学会。
A robust and sensitive sample preparation method is presented for matrix-assisted laser desorption ionization (MALDI) mass spectrometric analysis of low nanomolar concentrations of proteins containing high amounts of common salts and buffers. This method involves the production of densely packed sub-micrometer matrix crystals by depositing a matrix solution on top of a matrix seed-layer prepared on a MALDI target. A sub-microliter aliquot of analyte solution is then directly added to the top of the matrix crystals to form a thin-layer. alpha-Cyano-4-hydroxycinnamic acid (4-HCCA) is used as matrix and demonstrated to give better performance than other commonly used matrices, such as 2,5-dihydroxybenzoic acid (DHB), 2-(4-hydroxy-phenylazo) benzoic acid (HABA), or sinapinic acid. This three-layer method is shown to be superior to the other MALDI sample preparation methods, particularly for handling low nanomolar protein solutions containing salts and buffers. (J Am Soc Mass Spectrom 2006, 17, 780-785) (C) 2006 American Society for Mass Spectrometry.