Improved intensity-based label-free quantification via proximity-based intensity normalization (PIN).

Improved intensity-based label-free quantification via proximity-based intensity normalization (PIN).
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DOI:
10.1021/pr400866r
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发表时间:
2014-03-07
影响因子:
4.4
通讯作者:
Griffin TJ
Griffin TJ
中科院分区:
生物学2区
文献类型:
--
作者:
Van Riper SK;de Jong EP;Higgins L;Carlis JV;Griffin TJ

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研究人员越来越多地在高效液相-电喷雾质谱/质谱仪工作流程中转向基于MS1强度的无标记量化策略,以揭示分子水平上的生物变异。不幸的是,使用这些策略的高效液相-电喷雾质谱/质谱仪工作流程产生的结果重复性和再现性较差,这主要是由于系统偏差和复杂的可变性。虽然目前的全球归一化策略可以减轻系统偏差,但在高效液相-电喷雾质谱/质谱仪分析过程中,当面临由瞬时随机事件引起的复杂变异性时,它们会失败。为了解决这些问题,我们在分析成分数据的基础上提出了一种新的局部归一化方法--基于邻近度的强度归一化(PIN)。我们对照常见的标准化策略对PIN进行了评估。PIN在极大地减少差异以及识别出20%以上具有统计意义的丰度差异的蛋白质方面优于它们,这是其他策略所错过的。我们的结果表明,PIN能够发现统计上意义重大的生物变异,否则就会被错误报告或遗漏。
Researchers are increasingly turning to label-free MS1 intensity-based quantification strategies within HPLC–ESI–MS/MS workflows to reveal biological variation at the molecule level. Unfortunately, HPLC–ESI–MS/MS workflows using these strategies produce results with poor repeatability and reproducibility, primarily due to systematic bias and complex variability. While current global normalization strategies can mitigate systematic bias, they fail when faced with complex variability stemming from transient stochastic events during HPLC–ESI–MS/MS analysis. To address these problems, we developed a novel local normalization method, proximity-based intensity normalization (PIN), based on the analysis of compositional data. We evaluated PIN against common normalization strategies. PIN outperforms them in dramatically reducing variance and in identifying 20% more proteins with statistically significant abundance differences that other strategies missed. Our results show the PIN enables the discovery of statistically significant biological variation that otherwise is falsely reported or missed.
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