Purification of contaminated peptides and proteins on synthetic membrane surfaces for matrix-assisted laser desorption/ionization mass spectrometry
Purification of contaminated peptides and proteins on synthetic membrane surfaces for matrix-assisted laser desorption/ionization mass spectrometry
复制标题
DOI：
10.1021/ac970969e
复制
发表时间：
1998-02-15
影响因子：
7.4
通讯作者：
Woods, AS
中科院分区：
文献类型：
--
作者：
Worrall, TA;Cotter, RJ;Woods, AS
对盐、洗涤剂、甘油污染的肽和蛋白质样品进行有效的MALDI- tof质谱分析，通过将样品装载到合成膜上，洗去污染物，加入基质，并直接从膜表面解吸样品来完成，该方法容易去除妨碍有效MALDI的污染物；从不锈钢表面去除多肽和蛋白质，避免了费力的进一步纯化和相关的样品损失，聚乙烯，聚丙烯，C8和C18表面都被证明能有效去除污染物，样品表面的扫描电子显微镜图像显示，聚乙烯和聚丙烯表面形成的基质晶体明显小于不锈钢，C8或C18表面。从聚乙烯和聚丙烯表面解吸产生的可重复性光谱比从不锈钢表面解吸的样品质量分辨率更高，分辨率的提高，结合产物离子光谱强度的降低，表明从聚乙烯和聚丙烯表面解吸时肽和蛋白质分子离子的内能比从不锈钢表面解吸时少，污染样品的MALDI可以通过直接的，在探针上纯化，导致更高的分辨率光谱比观察到的样品从不锈钢解吸。
Effective MALDI-TOF MS of salt-, detergent-, and glycerol-contaminated peptide and protein samples is accomplished by loading samples onto synthetic membranes, washing away contaminants, adding matrix, and desorbing samples directly from the membrane surface, The method easily removes contaminants which prevent effective MALDI;DI of peptides and proteins from stainless steel surfaces, obviating the need for laborious further purification and associated sample losses, Polyethylene, polypropylene, C8, and C18 surfaces all proved effective at removing contaminants, Scanning electron microscope images of sample surfaces reveal that significantly smaller matrix crystals form on polyethylene and polypropylene, surfaces than on stainless steel, C8, or C18 surfaces. Desorption from polyethylene and polypropylene surfaces generates consistently reproducible spectra with better mass resolution than observed for samples desorbed from stainless steel, Improved resolution, combined with reduced intensity of product ion spectra, indicate that peptide and protein molecular ions have less internal energy when desorbed from polyethylene and polypropylene surfaces than from stainless steel, MALDI of contaminated samples can be accomplished by straightforward, on-probe purification, resulting in higher resolution spectra than observed in samples desorbed from stainless steel.