a Biotechnology Division , Biomedical Research Foundation, Academy of Athens (BRFAA) , Athens , Greece.
Expert Rev Mol Diagn. 2019 Jun;19(6):499-515. doi: 10.1080/14737159.2019.1615448. Epub 2019 May 17.
: Multiple (or selected) reaction monitoring-mass spectrometry (MRM/SRM) is a targeted proteomic method that can be used for relative and absolute quantification. Multiple reports exist supporting the potential of the approach in proteomic biomarker validation. : To get an overview of the applications of MRM in protein quantification in plasma, a search in MedLine/PubMed was performed using the keywords: 'MRM/SRM plasma proteomic/proteomics/proteome'. The retrieved studies were further filtered to focus on disease biomarkers and the main results are summarized. : MRM is increasingly employed for the quantification of both well-established but also newly discovered putative biomarkers and occasionally their post-translationally modified forms in plasma. Fractionation is regularly required for the detection of low abundance proteins. Standardized procedures to facilitate assay establishment and marker quantification have been proposed and, in few cases, implemented. Nevertheless, in most cases, absolute quantification is not performed. To advance, multiple technical issues including the regular use of standard labeled peptides and appropriate quality controls to monitor assay performance should be considered. Additionally, clinical aspects involving careful study design to address biomarker clinical use should also be considered.
多反应监测/选择反应监测-质谱(MRM/SRM)是一种靶向蛋白质组学方法,可用于相对和绝对定量。有多个报告支持该方法在蛋白质组学生物标志物验证中的潜力。为了全面了解 MRM 在血浆蛋白质定量中的应用,在 MedLine/PubMed 中使用关键词“MRM/SRM 血浆蛋白质组学/蛋白质组学/蛋白质组”进行了搜索。进一步筛选检索到的研究,重点关注疾病生物标志物,并总结主要结果。MRM 越来越多地用于定量分析已建立的但也有新发现的潜在生物标志物及其在血浆中的翻译后修饰形式。为了检测低丰度蛋白质,通常需要进行分级分离。已经提出并在少数情况下实施了标准化程序来促进检测方法的建立和标志物的定量。然而,在大多数情况下,没有进行绝对定量。为了取得进展,应考虑多个技术问题,包括经常使用标准标记肽和适当的质量控制来监测检测性能。此外,还应考虑涉及仔细研究设计以解决生物标志物临床应用的临床方面。