Protein identification at the low femtomole level from silver-stained gels using a new fritless electrospray interface for liquid chromatography microspray and nanospray mass spectrometry
Protein identification at the low femtomole level from silver-stained gels using a new fritless electrospray interface for liquid chromatography microspray and nanospray mass spectrometry
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DOI：
10.1006/abio.1998.2809
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发表时间：
1998-10-01
影响因子：
2.9
通讯作者：
Yates, JR
中科院分区：
文献类型：
--
作者：
Gatlin, CL;Kleemann, GR;Yates, JR
常规的毛细管液相色谱/质谱法（LC/MS）通常采用低μ l/min流速和气/液鞘以增强喷雾稳定性。在过去的几年中，许多报告已经证明了成功的电喷雾（ES）接口设计优化亚微升/分钟的方式，具有明显的优势，在提高检测限，降低样品消耗，并能够适应更广泛的缓冲条件。我们在这里报告的无熔块电喷雾接口（FESI）的设计，是廉价和强大的，可以操作和适应各种应用亚微升/分钟的流速。该接口的新奇之处在于使用无熔块微毛细管柱并在微三通接头处在柱前施加电喷雾电压，以实现稳定的微喷雾和纳米喷雾流速。这种无鞘FESI装置消除了柱后死体积，因为小颗粒（小于或等于10 μ m）被直接填充到激光牵引的熔融石英毛细管针中，从那里产生喷雾。对于溶液中的蛋白质/肽的分析，已经实现了低飞摩尔灵敏度（阿托摩尔用于选择离子监测），而对于源自来自1-D和2-D凝胶的凝胶内消化的银染色条带的蛋白质，实现了低纳克灵敏度。提出了使用LC-微喷雾、LC-纳米喷雾或输注纳米喷雾的串联MS蛋白质/肽鉴定的几种应用，（C）1998 Academic Press。
Conventional capillary liquid chromatography/mass spectrometry (LC/MS) typically employs low mu l/min how rates with gas/liquid sheath to enhance spray stability. Over the past several years a number of reports have demonstrated success with electrospray (ES) interface designs optimized for submicroliter/min hows which have clear advantages in terms of enhancement of detection Limit, lower sample consumption, and ability to accommodate a wider range of buffer conditions. We report here a fritless electrospray inter-face (FESI) design that is inexpensive and robust and can be operated and adapted to accommodate a variety of applications for submicroliter/min flow rates. The novelty of this interface revolves around the use of a fritless microcapillary column and precolumn application of electrospray voltage at a microtee junction to achieve stable microspray and nanospray flow rates. This sheathless FESI device eliminates postcolumn dead volume since small particles (less than or equal to 10 mu m) are packed directly into laser-pulled fused silica capillary needles from which a spray originates. For analysis of proteins/peptides in solution, low femtomole sensitivity has been achieved (attomoles for selected-ion monitoring), while low nanogram sensitivity was attained for proteins derived from in-gel-digested silver-stained bands from 1-D and 2-D gels, Several applications for tandem MS protein/peptide identification using LC-microspray, LC-nanospray, or infusion nanospray are presented, (C) 1998 Academic Press.