Capillary isoelectric focusing coupled offline to matrix assisted laser desorption/ionization mass spectrometry
Capillary isoelectric focusing coupled offline to matrix assisted laser desorption/ionization mass spectrometry
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DOI：
10.1016/j.chroma.2009.11.047
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发表时间：
2010-01-01
影响因子：
4.1
通讯作者：
Hayes, Mark A.
中科院分区：
文献类型：
--
作者：
Weiss, Noah G.;Zwick, Nicole L.;Hayes, Mark A.
这项工作提出了几个关键的细节，使cIEF-MALDI-MS一个强大的技术，这将允许更多的常规应用和自动化的援助。这包括强调注射泵移动所需的硬件和防止气泡破坏的适当方案。根据这些指导原则，6个pl市场的洗脱时间重现性极佳（RSD < 5%）。此外，pI市场用于校准pH梯度并确定通过质谱法离线检测的蛋白质的实验pI。这是用肌红蛋白和两种形式的β-乳球蛋白的标准蛋白质混合物证明的。实验测定的蛋白质的PLS和分子量被发现与文献值一致。所讨论的技术细节为应用MALDI-MS与cIEF的离线耦合提供了良好的基础。(C)2009 Elsevier B. V.保留所有权利。
This work presents several critical details for making cIEF-MALDI-MS a robust technique which will allow for more routine application and aid in automation. This includes emphasis on the hardware necessary for syringe pump mobilization and proper protocol for preventing disruption from gas bubbles. Following these guidelines, excellent elution time reproducibility is demonstrated for six pl markets (RSD < 5%). Additionally, the pl markets are used to calibrate the pH gradient and determine experimental pis of proteins detected offline by mass spectrometry. This was demonstrated using a standard protein mixture of myoglobin and two forms of beta-lactoglobulin. Experimental determination of protein pls and molecular weights were found to be in agreement with literature values. The technical details discussed provide a sound foundation for applying the offline coupling of MALDI-MS with cIEF. (C) 2009 Elsevier B.V. All rights reserved.