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、预后和治疗方面具有巨大的潜力。实现癌症生物标志物的高治疗和金融潜力需要新的技术来快速和经济地生产高质量的生物标志物，并对新生产的生物标志物进行方便的调整。候选生物标志物特征肽的液相色谱-稳定同位素稀释-多重反应监测质谱（LC-SID-MRM MS）是一种非常适合该领域的新兴技术。本应用将确立转化该质谱方法突破样品通量瓶颈的可行性。这项新技术被称为超通量多反应监测（UMRM）质谱，是lc - sidd - mrm质谱和肽衍生化的集成。两个特定的目标将被追求：(1)鉴定适合UMRM测量的肽衍生化，以及(2)在单个实验中验证衍生化用于108个血清样本中前列腺特异性抗原的UMRM分析。新技术的基本新颖性在于LC-SID-MRM MS的新转变，以产生前所未有的样品吞吐量。它有三个独特的优势：吞吐量优势，灵活性和即时性优势，以及经济优势。这项新技术可以通过廉价的试剂和商用质谱仪来实现，在生物标志物管道的许多阶段对癌症生物标志物候选物进行集中定量。新技术大幅增加的样品吞吐量将特别影响生物标志物候选物的验证和验证，这需要对数百到数千个患者对照样品进行靶向定量。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. PUBLIC HEALTH RELEVANCE: This project examines the feasibility of transforming he multiplexing potential to the sample- throughput potential of the emerging peptide multiple reaction monitoring mass spectrometry for cancer biomarker development. The new technology, capable of one-experiment quantitation of large numbers of samples, will significantly impact the large-scale validation of cancer biomarkers which uses hundreds and thousands of patient-control samples.