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首页> 外文期刊>Analytical chemistry >Qualitative and Quantitative Characterization of Plasma Proteins When Incorporating Traveling Wave Ion Mobility into a Liquid Chromatography-Mass Spectrometry Workflow for Biomarker Discovery: Use of Product Ion Quantitation As an Alternative Data Analys is Tool for Label Free Quantitation
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Qualitative and Quantitative Characterization of Plasma Proteins When Incorporating Traveling Wave Ion Mobility into a Liquid Chromatography-Mass Spectrometry Workflow for Biomarker Discovery: Use of Product Ion Quantitation As an Alternative Data Analys is Tool for Label Free Quantitation

机译：将行波离子迁移率纳入液相色谱-质谱分析工作流程以进行生物标记物发现时，血浆蛋白的定性和定量表征：使用产物离子定量作为替代数据分析是无标签定量的工具

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Discovery of protein biomarkers in clinical samples necessitates significant prefractionation prior to liquid chromatography-mass spectrometry (LC-MS) analysis. Integrating traveling wave ion mobility spectrometry (TWIMS) enables in-line gas phase separation which when coupled with nanoflow liquid chromatography and data independent acquisition tandem mass spectrometry, confers significant advantages to the discovery of protein biomarkers by improving separation and inherent sensitivity. Incorporation of TWIMS leads to a packet of concentrated ions which ultimately provides a significant improvement in sensitivity. As a consequence of ion packeting, when present at high concentrations, accurate quantitation of proteins can be affected due to detector saturation effects. Human plasma was analyzed in triplicate using liquid-chromatography data independent acquisition mass spectrometry (LC-DIA-MS) and using liquid-chromatography ion-mobility data independent acquisition mass spectrometry (LC-IM-DIA-MS). The inclusion of TWIMS was assessed for the effect on sample throughput, data integrity, confidence of protein and peptide identification, and dynamic range. The number of identified proteins is significantly increased by an average of 84% while both the precursor and product mass accuracies are maintained between the modalities. Sample dynamic range is also maintained while quantitation is achieved for all but the most abundant proteins by incorporating a novel data interpretation method that allows accurate quantitation to occur. This additional separation is all achieved within a workflow with no discernible deleterious effect on throughput. Consequently, TWIMS greatly enhances proteome coverage and can be reliably used for quantification when using an alternative product ion quantification strategy. Using TWIMS in biomarker discovery in human plasma is thus recommended.
机译：在临床样品中发现蛋白质生物标志物需要在液相色谱-质谱（LC-MS）分析之前进行大量的预分离。集成行波离子迁移谱（TWIMS）可以实现在线气相分离，当与纳流液相色谱和数据独立采集串联质谱联用时，通过改进分离和固有灵敏度，可以为发现蛋白质生物标记物带来显着优势。掺入TWIMS会导致一包浓缩离子，最终使灵敏度大大提高。作为离子包封的结果，当以高浓度存在时，由于检测器饱和效应，可能会影响蛋白质的准确定量。使用液相色谱数据独立采集质谱仪（LC-DIA-MS）和液相色谱离子迁移率数据独立采集质谱仪（LC-IM-DIA-MS）一式三份分析人血浆。评估了TWIMS的加入对样品通量，数据完整性，蛋白质和多肽鉴定的可信度以及动态范围的影响。鉴定出的蛋白质数量平均显着增加了84％，同时在这些模式之间保持了前体和产物质量的准确性。通过结合允许精确定量发生的新颖数据解释方法，在维持除最丰富蛋白质外的所有蛋白质定量的同时，还可以保持样品动态范围。所有这些额外的分离都是在工作流程中实现的，并且对吞吐量没有明显的有害影响。因此，TWIMS大大提高了蛋白质组覆盖率，并且在使用替代产物离子定量策略时可以可靠地用于定量。因此建议在人血浆中的生物标志物发现中使用TWIMS。

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