College of Food Science and Nutritional Engineering , China Agricultural University , No. 17 Qinghua East Road , Haidian District, Beijing 100083 , P. R. China.
Department of Molecular Biology and Genetics, Faculty of Science and Technology , Aarhus University , Gustav Wieds Vej 10 , 8000 Aarhus , Denmark.
J Agric Food Chem. 2019 Feb 6;67(5):1608-1618. doi: 10.1021/acs.jafc.8b05459. Epub 2019 Jan 23.
Mass spectrometry (MS) has become essential for efficient and accurate quantification of proteins and proteomes and, thus, a key technology throughout all biosciences. However, validated MS methods are still scarce for meat quality research applications. The objective of this work was to develop and compare two targeted proteomic methods, namely, selected reaction monitoring (SRM) and sequential window acquisition of all theoretical spectra (SWATH), for the quantification of 11 bovine muscle proteins that may be indicators of meat color. Both methods require evaluation of spectra from proteotypic and quantotypic peptides, and we here report our evaluation of which peptides and MS parameters are best suited for robust quantification of these 11 proteins. We observed that the SRM approach provides better reproducibility, linearity, and sensitivity than SWATH and is therefore ideal for targeted quantification of low-abundance proteins, while the SWATH approach provides a more time-efficient method for targeted protein quantification of high-abundance proteins and, additionally, supports the search for novel biomarkers.
质谱（MS）已成为高效、准确地定量蛋白质和蛋白质组的必要手段，因此成为所有生物科学的关键技术。然而，经过验证的 MS 方法仍然在肉类质量研究应用中相对较少。本研究的目的是开发和比较两种靶向蛋白质组学方法，即选择反应监测（SRM）和全理论谱序贯窗口采集（SWATH），用于定量 11 种可能作为肉色指标的牛肌肉蛋白。这两种方法都需要对特征肽和定量肽的谱进行评估，我们在这里报告了我们对哪些肽和 MS 参数最适合于这些 11 种蛋白质的稳健定量的评估。我们观察到，SRM 方法比 SWATH 具有更好的重现性、线性和灵敏度，因此非常适合低丰度蛋白质的靶向定量，而 SWATH 方法则提供了一种更高效的靶向定量方法，用于高丰度蛋白质的定量，此外，还支持寻找新的生物标志物。
