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定量蛋白质组学质谱采集技术进展（四）

2020.5.18

无论是相对定量还是绝对定量方法,DIA很好地克服了DDA鸟枪法和SRM目标监测的种种不足, 在定量蛋白质组学中具有良好的应用前景。然而,目前DIA 方法的循环时间仍然较长,只能与纳流液相联用,并使用较长的梯度以获得足够的色谱峰宽,限制了DIA 的应用范围。这也是DIA 技术下一步需要解决的问题。

图7WiSIM-DIA 与Full MS-DIA 原理[57] :(A) WiSIM-DIA 采集原理; (B) Full MS-DIA 采集原理

Fig. 7Schematic elucidation of wide isolation-SIM (WiSIM)-DIA and Full MS-DIA[57] : (A) Workflow of WiSIM-DIA; (B) Workflow of Full MS-DIA

5 总结与展望

基于报告离子定量的同重同位素标记、目标离子监测和数据非依赖采集已成为定量蛋白质组学的主要技术手段,表1 总结和比较了这3 种方法的原理和异同。定量蛋白质组学的飞速发展为质谱技术带来挑战,基于稳定同位素标记的相对定量和基于SRM 的绝对定量都面临着复杂基质的严重干扰和通量不足等局限(表1)。而近来一系列高分辨质谱新技术的发展为解决这些问题带来希望。其中,同步母离子选择和质量亏损标记有效解决了相对定量的干扰和通量问题; 平行反应监测及多重累积技术提高了SRM 的选择性,成为绝对定量的新途径; 数据非依赖性采集兼具DDA 与SRM 的优势,多重累积和三合一质谱技术使DIA 的扫描步长进一步缩小,能更有效地使DIA 技术应用于高通量的定量蛋白质组学。未来,这些新技术将逐渐取代传统质谱技术,越来越多地应用到定量蛋白质组学中,为解决诸如蛋白质相互作用、临床标志物研究等领域最棘手的问题带来新的手段和突破。

表1 定量蛋白质组学主要技术的原理与进展

Table 1 Principle and progress of quantitative proteomic methods

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Progress in Mass Spectrometry Acquisition Approach for Quantitative Proteomics

ZHANG Wei*

(Thermo Fisher Scientific, Shanghai 201206, China)

Abstract Mass spectrometry is an important and powerful tool for protein quantification. With the in -depth development of quantitative proteomics, limitations of classic MS based quantification methods, such as complicated matrix interference and throughput/ capacity limitation, start to appear. Recent progress of series novel MS based techniques provide effective solutions for the limitations of relative and absolute proteomic quantification, including synchronous precursor selection ( SPS), mass defect isobaric labeling, parallel reaction monitoring (PRM), multiplexing acquisition (MSX), and various novel data independent acquisition(DIA) modes. Here we summarized the current limitations of quantitative proteomics, reviewed the latest MS based quantification approaches, and discussed the features and advantages of these novel techniques for quantitative proteomic application.

Keywords Quantitative proteomics; Synchronous precursor selection; Parallel reaction monitoring; Data independent acquisition; Review

(Received 10 September 2014; accepted 18 October 2014)

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