Liquid ultraviolet matrix-assisted laser desorption/ionization mass spectrometry for automated proteomic analysis
Liquid ultraviolet matrix-assisted laser desorption/ionization mass spectrometry for automated proteomic analysis
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DOI：
10.1002/pmic.200400956
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发表时间：
2005-02-01
期刊：
影响因子：
3.4
通讯作者：
Corless, S
中科院分区：
文献类型：
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作者：
Cramer, R;Corless, S
我们结合了几个关键的样品制备步骤，以使用液体基质系统，在自动紫外基质辅助激光解吸/电离-质谱仪(UV-MALDI-MS)中提供高分析灵敏度。这种新的样品制备方法采用了一种基于Sze等人描述的甘油基质混合物的基质混合物。美国联合航空公司SoC。质谱学。1998、9、166-174)。这种新的制备方法可以实现低的蛋白质摩尔灵敏度，使得蛋白质消化的蛋白质组学分析可以与固态基质系统相媲美。在自动数据采集和分析方面，这种液体基质的MALDI性能超过了传统的固态MALDI矩阵。除了液体样品用于自动样品制备和数据采集的固有一般优势外，与通常使用的固体基质(如2，5-二羟基苯甲酸(DHB)或α-氰基-羟基肉桂酸(CHCA))相比，使用所提供的液体基质显著减少了多肽质量指纹图谱中非特定离子信号的范围。特别是，基质和低质量离子信号以及由阳离子加合物形成产生的离子信号显著减少。因此，通过溶液内和凝胶内消化的肽质量图谱进行蛋白质鉴定的置信度通常更高。
We have combined several key sample preparation steps for the use of a liquid matrix system to provide high analytical sensitivity in automated ultraviolet - matrix-assisted laser desorption/ ionisation - mass spectrometry (UV-MALDI-MS). This new sample preparation protocol employs a matrix-mixture which is based on the glycerol matrix-mixture described by Sze et al. U. Am. Soc. Mass Spectrom. 1998, 9, 166-174). The low-ferntomole sensitivity that is achievable with this new preparation protocol enables proteomic analysis of protein digests comparable to solid-state matrix systems. For automated data acquisition and analysis, the MALDI performance of this liquid matrix surpasses the conventional solid-state MALDI matrices. Besides the inherent general advantages of liquid samples for automated sample preparation and data acquisition the use of the presented liquid matrix significantly reduces the extent of unspecific ion signals in peptide mass fingerprints compared to typically used solid matrices, such as 2,5-dihydrox-ybenzoic acid (DHB) or alpha-cyano-hydroxycinnamic acid (CHCA). In particular, matrix and lowmass ion signals and ion signals resulting from cation adduct formation are dramatically reduced. Consequently, the confidence level of protein identification by peptide mass mapping of in-solution and in-gel digests is generally higher.