Ultrathroughput Multiple Reaction Monitoring Mass Spectrometry for Large-Scale Ca

描述（由申请人提供）：蛋白质生物标志物在癌症的诊断、预后和治疗中具有巨大的潜力。实现癌症生物标志物的高治疗和经济潜力需要新技术来快速和具有成本效益地生产高质量的生物标志物，以及用于测量新产生的生物标志物的容易的调整。候选生物标志物的特征肽的液相色谱-稳定同位素稀释-多反应监测质谱（LC-SID-MRM MS）是非常适合该领域的新兴技术。该应用将确立改造这种质谱方法以突破样品通量瓶颈的可行性。新技术被称为超通量多反应监测（UMRM）MS，是LC-SID-MRM MS和肽衍生化的集成。将追求两个具体目标：（1）鉴定用于UMRM测量的合适肽衍生化，和（2）在单个实验中验证用于108份血清样品中前列腺特异性抗原的UMRM分析的衍生化。新技术的基本新奇在于LC-SID-MRM MS的新颖转换，以产生前所未有的样品通量。它具有三个独特的优势：吞吐量优势、灵活性和即时性优势以及经济性优势。这项新技术可以用廉价的试剂和商业质谱仪实现，用于在生物标志物管道的许多阶段集中定量癌症生物标志物候选物。新技术大幅增加的样品通量将特别影响生物标志物候选物的验证和确认，这需要对数百至数千个患者对照样品进行有针对性的定量。因此，UMRM技术将显著加速新的癌症生物标志物的产生。这项新技术在推进癌症的定量生物学方面也有潜力。

DESCRIPTION (provided by applicant): Protein biomarkers have enormous potential in the diagnosis, prognosis and treatment of cancer. Realization of the high therapeutic and financial potential of cancer biomarkers demands new technologies for speedy and cost-effective production of high-quality biomarkers and facile adjustments for measuring newly-produced biomarkers. Liquid chromatography- stable isotope dilution-multiple reaction monitoring mass spectrometry (LC-SID-MRM MS) of signature peptides of candidate biomarkers is an emerging technology well suited to this field. This application will establish the feasibility of transforming this mass spectrometry method to break through the sample-throughput bottleneck. The new technology is termed Ultrathroughput Multiple Reaction Monitoring (UMRM) MS and is an integration of LC-SID-MRM MS and peptide derivatization. Two specific aims will be pursued: (1) identification of suitable peptide derivatizations for UMRM measurements, and (2) validation of derivatizations for UMRM analysis of prostate-specific antigen in 108 serum samples in a single experiment. The fundamental novelty of the new technology rests on the novel transformation of LC-SID-MRM MS to produce unprecedented sample throughput. It has three unique advantages: the throughput advantage, the flexibility and immediate-impact advantage, and the economic advantage. The new technology can be implemented with inexpensive reagents and commercial mass spectrometers for focused quantitation of cancer biomarker candidates in many phases of a biomarker pipeline. The drastically increased sample throughput of the new technology will in particular impact the verification and validation of biomarker candidates, which require targeted quantitation of hundreds to thousands of patient-control samples. Thus, the UMRM technology will significantly accelerate new cancer biomarker generation. The new technology also has potential in advancing quantitative biology of cancer.