Ultrathroughput Multiple Reaction Monitoring Mass Spectrometry for Large-Scale Ca

描述(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。实现癌症生物标记物的高治疗和经济潜力需要新的技术，以快速和具有成本效益的方式生产高质量的生物标记物，并对新生产的生物标记物进行简便的调整。候选生物标志物标志肽的LC-SID-MRM-MS是一项非常适合这一领域的新兴技术。这一应用将确立改变这种质谱学方法以突破样品吞吐瓶颈的可行性。这项新技术被称为超通量多反应监测(UMRM)MS，是LC-SID-MRM MS和多肽衍生化的集成。将追求两个具体目标：(1)为UMRM测量确定合适的多肽衍生物；(2)在一次实验中对108个血清样本的UMRM前列腺特异性抗原的衍生物进行验证。这项新技术的根本创新之处在于对LC-SID-MRM MS进行了新的改造，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、灵活性和立竿见影的优势、经济优势。这项新技术可以与廉价的试剂和商业质谱仪一起实施，以便在生物标记物管道的许多阶段集中定量癌症生物标记物候选。新技术大幅增加的样本产量将特别影响生物标记物候选的验证和确认，这需要对数百到数千个患者对照样本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记物的产生。这项新技术在推进癌症定量生物学方面也具有潜力。 公共卫生相关性：该项目研究了将多路传输潜力转化为新出现的多肽多反应监测质谱学用于癌症生物标志物开发的样品吞吐潜力的可行性。这项新技术能够对大量样本进行一次实验量化，将对使用成百上千个患者对照样本的癌症生物标记物的大规模验证产生重大影响。

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.