Ibrahim Yehia M, Belov Mikhail E, Liyu Andrei V, Smith Richard D
Biological Sciences Division, Pacific Northwest National Laboratory, P.O. Box 999, Richland, Washington 99352, USA.
Anal Chem. 2008 Jul 15;80(14):5367-76. doi: 10.1021/ac8003488. Epub 2008 May 31.
Time-of-flight mass spectrometry (TOF MS) is increasingly used in proteomics research. Herein, we report on the development and characterization of a TOF MS instrument with improved sensitivity equipped with an electrodynamic ion funnel trap (IFT) that employs an automated gain control (AGC) capability. The IFT-TOF MS was coupled to a reversed-phase capillary liquid chromatography (RPLC) separation and evaluated in experiments with complex proteolytic digests. When applied to a global tryptic digest of Shewanella oneidensis proteins, an order-of-magnitude increase in sensitivity compared to that of the conventional continuous mode of operation was achieved due to efficient ion accumulation prior to TOF MS analysis. As a result of this sensitivity improvement and related improvement in mass measurement accuracy, the number of unique peptides identified in the AGC-IFT mode was 5-fold greater than that obtained in the continuous mode.
飞行时间质谱（TOF MS）在蛋白质组学研究中的应用越来越广泛。在此，我们报告了一种配备电动离子漏斗阱（IFT）且具有自动增益控制（AGC）功能的灵敏度更高的TOF MS仪器的开发与特性。该IFT-TOF MS与反相毛细管液相色谱（RPLC）分离联用，并在复杂蛋白酶解消化实验中进行了评估。当应用于嗜冷栖热放线菌蛋白质的全局胰蛋白酶消化时，由于在TOF MS分析之前进行了有效的离子积累，与传统连续操作模式相比，灵敏度提高了一个数量级。由于灵敏度的提高以及质量测量精度的相关提高，在AGC-IFT模式下鉴定出的独特肽段数量比连续模式下多5倍。
