Dual-pressure linear ion trap mass spectrometer improving the analysis of complex protein mixtures.
Dual-pressure linear ion trap mass spectrometer improving the analysis of complex protein mixtures.
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DOI：
10.1021/ac901278y
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发表时间：
2009-09-15
影响因子：
7.4
通讯作者：
Zabrouskov, Vlad
中科院分区：
文献类型：
--
作者：
Second, Tonya Pekar;Blethrow, Justin D.;Schwartz, Jae C.;Merrihew, Gennifer E.;MacCoss, Michael J.;Swaney, Danielle L.;Russell, Jason D.;Coon, Joshua J.;Zabrouskov, Vlad
在过去的十年中，通过液相色谱-电喷雾电离-串联质谱技术进行高通量蛋白质组学分析取得了相当大的进展，这在很大程度上是由于仪器的不断改进。高通量鉴定实验以肽测序为基础，主要通过串联质谱法完成，离子阱和基于阱的仪器已成为广泛采用的分析平台。为了满足对表征深度和通量日益苛刻的要求，我们提出了一种新开发的双压力线性离子阱质谱仪（LTQ Velos），该质谱仪具有更高的灵敏度，提供了新的源设计，并且实际循环时间比LTQ XL短两倍，同时提高或保持了MS/MS碎片光谱的光谱质量。与现有平台相比，这些改进导致了秀丽隐杆线虫复杂蛋白质组中蛋白质和独特肽的检测和鉴定的大幅增加。进入质谱仪的离子通量大大增加，再加上对低丰度前体离子的分离改进，导致低丰度肽的检测增加。与LTQ XL相比，这些改进累积起来导致了对面包酵母（酿酒酵母）蛋白质组的更大渗透。另外，新仪器上更快的循环时间允许对给定深度的蛋白质组分析提供更高的通量，使用LTQ Velos在60分钟内识别更多的肽和蛋白质，而使用LTQ XL在180分钟内识别更多的肽和蛋白质。当质量分析以超过25,000 FWHM的分辨率进行时，可以同位素解析小的完整蛋白质及其片段，从而为自上而下的实验打开了可能性。
The considerable progress in high throughput proteomics analysis via liquid chromatography-electrospray ionization-tandem mass spectrometry over the last decade has been fueled to a large degree by continuous improvements in instrumentation. High throughput identification experiments are based on peptide sequencing and are largely accomplished through the use of tandem mass spectrometry, with ion trap and trap-based instruments having become broadly adopted analytical platforms. To satisfy increasingly demanding requirements for depth of characterization and throughput, we present a newly developed dual-pressure linear ion trap mass spectrometer (LTQ Velos) that features increased sensitivity, afforded by a new source design, and demonstrates practical cycle times two times shorter than that of an LTQ XL, while improving or maintaining spectral quality for MS/MS fragmentation spectra. These improvements resulted in a substantial increase in the detection and identification of both proteins and unique peptides from the complex proteome of Caenorhabditis elegans, as compared to existing platforms. The greatly increased ion flux into the mass spectrometer in combination with improved isolation of low-abundance precursor ions resulted in increased detection of low-abundance peptides. These improvements cumulatively resulted in a substantially greater penetration into the baker’s yeast (Saccharomyces cerevisiae) proteome compared to LTQ XL. Alternatively, faster cycle times on the new instrument allowed for higher throughput for a given depth of proteome analysis, with more peptides and proteins identified in 60 min using an LTQ Velos than in 180 min using an LTQ XL. When mass analysis was carried out with resolution in excess of 25,000 FWHM, it became possible to isotopically resolve a small intact protein and its fragments, opening possibilities for top down experiments.
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影响因子：
7
作者：
Merrihew, Gennifer E.;Davis, Colleen;MacCoss, Michael J.
通讯作者：
MacCoss, Michael J.
DOI：
10.1073/pnas.172183199
发表时间：
2002-09-03
影响因子：
11.1
作者：
Koller, A;Washburn, MP;Yates, JR
通讯作者：
Yates, JR
影响因子：
2
作者：
McDonald, WH;Tabb, DL;Yates, JR
通讯作者：
Yates, JR
DOI：
10.1073/pnas.1530509100
发表时间：
2003-08-05
影响因子：
11.1
作者：
Storey, JD;Tibshirani, R
通讯作者：
Tibshirani, R
影响因子：
6.6
作者：
Kelly, Ryan T.;Tolmachev, Aleksey V.;Page, Jason S.;Tang, Keqi;Smith, Richard D.
通讯作者：
Smith, Richard D.