Evaluation of direct infusion-multiple reaction monitoring mass spectrometry for quantification of heat shock proteins.
Evaluation of direct infusion-multiple reaction monitoring mass spectrometry for quantification of heat shock proteins.
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DOI：
10.1021/ac203011j
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发表时间：
2012-02-21
影响因子：
7.4
通讯作者：
Koomen, John M.
中科院分区：
文献类型：
--
作者：
Xiang, Yun;Koomen, John M.
用液相色谱-多重反应监测质谱法（LC-MRM）进行蛋白质定量已经成为评估生物标志物的一个强大平台。在这项研究中，直接输注，使用自动化的，基于芯片的纳米电喷雾电离，结合MRM （DI-MRM）用于蛋白质定量。移除LC分离步骤增加了评估过渡之间比率的重要性。因此，溶剂组成、分析物浓度、喷雾电压和四极杆分辨率设置对碎片模式的影响已经使用肽和蛋白质标准进行了研究。在DI-MRM定量评估为标准后，热休克蛋白（HSPs）表达的定量分析从LC-MRM翻译为DI-MRM，用于多发性骨髓瘤细胞系模型。描述了DI-MRM分析开发的要求。然后，对两种方法进行了比较；有效的DI-MRM分析标准是基于分析全细胞裂解物中热休克蛋白的表达。高通量的DI-MRM分析有助于大批量类似样品的快速分析，如细胞对治疗反应的时间过程测量。
Protein quantification with liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM) has emerged as a powerful platform for assessing panels of biomarkers. In this study, direct infusion, using automated, chip-based nanoelectrospray ionization, coupled with MRM (DI-MRM) is used for protein quantification. Removal of the LC separation step increases the importance of evaluating the ratios between the transitions. Therefore, the effects of solvent composition, analyte concentration, spray voltage, and quadrupole resolution settings on fragmentation patterns have been studied using peptide and protein standards. After DI-MRM quantification was evaluated for standards, quantitative assays for the expression of heat shock proteins (HSPs) were translated from LC-MRM to DI-MRM for implementation in cell line models of multiple myeloma. Requirements for DI-MRM assay development are described. Then, the two methods are compared; criteria for effective DI-MRM analysis are reported based on the analysis of HSP expression in digests of whole cell lysates. The increased throughput of DI-MRM analysis is useful for rapid analysis of large batches of similar samples, such as time course measurements of cellular responses to therapy.
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DOI：
10.1016/j.jasms.2009.05.014
发表时间：
2009-09
影响因子：
3.2
作者：
Ferguson JT;Wenger CD;Metcalf WW;Kelleher NL
通讯作者：
Kelleher NL
DOI：
10.1074/mcp.m110.001537
发表时间：
2010-10
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Geiger T;Cox J;Mann M
通讯作者：
Mann M
影响因子：
7.4
作者：
Barnidge, DR;Dratz, EA;Lindall, A
通讯作者：
Lindall, A
影响因子：
4.8
作者：
Kirkpatrick, DS;Gerber, SA;Gygi, SP
通讯作者：
Gygi, SP
影响因子：
7
作者：
Kuzyk, Michael A.;Smith, Derek;Borchers, Christoph H.
通讯作者：
Borchers, Christoph H.