Lau Thomas Y K, Collins Ben C, Stone Peter, Tang Ning, Gallagher William M, Pennington Stephen R
UCD School of Biomolecular and Biomedical Science and Proteome Research Centre, UCD Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
Methods Mol Biol. 2011;691:417-27. doi: 10.1007/978-1-60761-849-2_25.
With the advent of "-omics" technologies, there has been an explosion of data generation in the field of toxicology, as well as in many others. As new candidate biomarkers of toxicity are being regularly discovered, the next challenge is to validate these observations in a targeted manner. Traditionally, these validation experiments have been conducted using antibody-based technologies such as Western blotting, ELISA, and immunohistochemistry. However, this often produces a significant bottleneck as the time, cost, and development of successful antibodies are often far outpaced by the generation of targets of interest. In response to this, recently there have been several developments in the use of triple quadrupole (QQQ) mass spectrometry (MS) as a platform to provide quantification of proteins by multiple reaction monitoring (MRM). This technology does not require antibodies; it is typically less expensive and quicker to develop, and has the opportunity for more accessible multiplexing. The speed of these experiments combined with their flexibility and ability to multiplex assays makes the technique a valuable strategy to validate biomarker discovery.
随着“组学”技术的出现，毒理学领域以及许多其他领域的数据生成量呈爆炸式增长。随着毒性新候选生物标志物不断被发现，下一个挑战是以有针对性的方式验证这些观察结果。传统上，这些验证实验是使用基于抗体的技术进行的，如蛋白质印迹法、酶联免疫吸附测定法和免疫组织化学法。然而，这往往会造成一个重大瓶颈，因为成功抗体的研发时间、成本与感兴趣靶标的产生速度相比往往远远滞后。对此，最近在使用三重四极杆（QQQ）质谱（MS）作为通过多反应监测（MRM）对蛋白质进行定量的平台方面有了一些进展。这项技术不需要抗体；它通常成本更低、开发更快，并且有机会实现更便捷的多重分析。这些实验的速度以及它们的灵活性和多重检测能力使该技术成为验证生物标志物发现的一种有价值的策略。
