使用商用微量馏分收集器进行板上消化，用于纳米高效液相色谱-基质辅助激光解吸/电离串联飞行时间蛋白质分析。
On-plate digestion using a commercial microfraction collector for nano-HPLC matrix-assisted laser desorption/ionization tandem time-of-flight protein analysis.
作者信息
Vitorino Rui, Guedes Sofia, Tomer Kenneth, Domingues Pedro, Duarte José, Amado Francisco
机构信息
Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.
出版信息
Anal Biochem. 2008 Sep 1;380(1):128-30. doi: 10.1016/j.ab.2008.05.006. Epub 2008 May 10.
Abstract
A new method for on-plate protein digestion and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry analysis is proposed involving an automated one-step sample separation using nanoflow HPLC followed by nanoliter fraction collection and on-plate digestion with trypsin. This procedure uses a commercial automatic nanoliter fraction collection system for on-line spotting of the eluent onto a MALDI target. After protein digestion, the reaction is stopped by the addition of acidified matrix using the same automated system. Collected spots are subsequently analyzed using a MALDI tandem time-of-flight (TOF/TOF) mass spectrometer for protein sequencing and identification.
摘要
提出了一种用于板上蛋白质消化和基质辅助激光解吸/电离（MALDI）质谱分析的新方法，该方法涉及使用纳流高效液相色谱（nanoflow HPLC）进行自动一步样品分离，随后进行纳升级分收集以及用胰蛋白酶进行板上消化。此程序使用商业自动纳升级分收集系统将洗脱液在线点样到MALDI靶板上。蛋白质消化后，使用同一自动系统加入酸化基质来终止反应。随后使用MALDI串联飞行时间（TOF/TOF）质谱仪对收集的斑点进行分析，以进行蛋白质测序和鉴定。
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