Validation and Advanced Development of Emerging Technologies

描述（由申请人提供）：该项目的主要目标是开发基于质谱（MS）的检测方法，用于高通量和定量生物标志物验证，其灵敏度和特异性与基于抗体的免疫检测方法相当或更好。该分析平台将主要基于三重四极杆质谱，通过整合最新的高速液相色谱（LC），太平洋西北国家实验室（PNNL）开发的高效电喷雾电离（ESI）源和独特的MS接口技术以及将仪器能力从执行传统的多反应监测（MRM）测量扩展到MRM 2 MS分析。基于新仪器的测定的预期定量限和动态范围预计将达到低pg/mL水平，并跨越样品浓度的八个数量级，从而允许定量测量复杂生物流体（如人血浆）中的低丰度蛋白质生物标志物。新的基于仪器的分析将适用于广泛的临床癌症生物标志物验证应用，并能够在一个实验中验证数百至数千个生物标志物。它们的性能将经过严格优化，并使用临床样本进行测试。

DESCRIPTION (provided by applicant): The primary goal of the project is to develop mass spectrometry (MS) based assays for high throughput and quantitative biomarker validation with sensitivity and specificity comparable to or better than antibody based immunoassays. The analytical platform will be primarily based on triple quadrupole mass spectrometry with its sensitivity, dynamic range, measurement specificity and sample throughput significantly enhanced by integrating the latest high speed liquid chromatography (LC), high efficiency electrospray ionization (ESI) source and unique MS interface technologies developed at Pacific Northwest National Laboratory (PNNL) and extending the instrument capability from performing traditional multiple reaction monitoring (MRM) measurements into MRM2 MS analysis. The expected limit of quantitation and dynamic range for the new instrument based assays will be expected to reach low pg/mL level and span eight orders of magnitude in sample concentration allowing quantitative measurements of low abundance protein biomarkers in complex biofluids, such as human blood plasma. The new instrument based assays will be applicable to a broad spectrum of clinical cancer biomarker validation applications and capable of validating hundreds to thousands biomarkers in a single experiment. Their performance will be rigorously optimized and tested using clinical samples.