Metal tagging of bioactive molecules for prognostic assays coupled with ICP-MS

描述（由申请人提供）：本提案的目的是开发一种先进的金属标记系统和方法，与电感耦合等离子体质谱仪（ICP-MS）检测器相结合，将为研究人员和临床医生提供大幅改善的分析和预后能力。ICP-MS作为分析检测器具有决定性的优势，可提高免疫测定的性能，包括：i）高精度; ii）低检测限; iii）大动态范围; iv）非特异性背景和分析响应与孵育或储存时间无关（因为蛋白质降解不影响元素标签的分析）; v）更大的多路复用潜力（潜在地，多达167种同位素;实际上，大约有100个可区分的标签。激发和吸收波长的失配以及发射信号的重叠是荧光团（包括量子点）的物理特性，其代表了在多重测定领域中进展的关键障碍并且不能被消除。我们的方法的一个关键特征是开发标记的亲和产物，其设计用于特异性识别区分细胞表面和细胞内标记物，并靶向用于ICP-MS检测。将开发用可区分的稳定同位素元素标签标记的方法。该项目将提供稳定的金属标签，适当标记的亲和试剂，方法和演示数据，用于区分多种分析物。我们希望这种集成的试剂系统将大大提高医生及其患者的诊断、预后和治疗效果，提高医疗保健的有效性，同时大幅降低现代个性化治疗的人力和财务成本。我们的跨学科专家小组将应对这一重大挑战。教授米切尔A. Winnik和Mark Nitz将利用现有能力来合成载体聚合物。将开发用可区分的稳定同位素元素标记进行标记的方法。ICP-MS组（弗拉基米尔I. Baranov）将开发一种特定目的的分析方法，将机器人样品引入与ICP-MS仪器相结合，从而实现多重检测。Olga I将开发元素标记的测定方法。Ornatskaia使用现有的能力和不断验证对传统的测定方法。

DESCRIPTION (provided by applicant): The objective of this proposal is development of an advanced metal tagging systems and methodology, which in combination with an Inductively Coupled Plasma Mass Spectrometer (ICP-MS) detector will provide researchers and clinicians with substantially improved analytical and prognostic capabilities. ICP-MS as an analytical detector possesses decisive advantages that enhance the performance of immunoassays, including: i) high precision; ii) low detection limits; iii) large dynamic range; iv) independence of non-specific background and analytical response from incubation or storage times (as protein degradation does not affect analysis of an elemental tag); v) larger multiplexing potential (potentially, up to 167 isotopes; realistically, around 100 distinguishable tags. Mismatch of excitation and absorption wavelengths, and overlap of emission signals are physical characteristics of fluorophores (including quantum dots), which represent a critical barrier to progress in the field of multiplexed assays and cannot be eliminated. A key feature of our approach is the development of tagged affinity products designed for specific recognition of distinguishing cell surface and intracellular markers and targeted for the ICP-MS detection. Methods of labeling with distinguishable stable isotopic elemental tags will be developed. This project will deliver stable metal tags, appropriately tagged affinity reagents, methods and demonstrative data for the distinction of multiple analytes. We hope that this integrated reagent system will dramatically enhance the diagnostic, prognostic and therapeutic efficacy available to physicians and their patients, increasing the effectiveness of healthcare while substantially reducing the human and financial costs of modern personalized treatment. Our interdisciplinary group of recognized experts will address this significant challenge. Professors Mitchell A. Winnik and Mark Nitz will use existing capabilities to synthesize the carrier polymers. Methods of tagging with distinguishable stable isotopic elemental tags will be developed. The ICP-MS group (Vladimir I. Baranov) will develop a purpose specific analytical methods combining robotic sample introduction with an ICP-MS instrument that will allow multiplex detection. Development of the elemental tagged assay methodology will be conducted by Olga I. Ornatskaia using existing capabilities and in constant verification against conventional assay methods.