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

描述(由申请人提供)：这项建议的目标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(ICP-MS)检测器相结合，将为研究人员和临床医生提供显著改善的分析和预测能力。作为一种分析检测器，电感耦合等离子体质谱具有提高免疫分析性能的决定性优势，包括：一)高精密度；二)低检测限；三)大动态范围；四)非特定背景和分析响应独立于孵育或储存时间(因为蛋白质降解不影响元素标签的分析)；五)更大的多重潜力(潜在地，多达167个同位素；实际上，大约100个可区分的标签)。激发波长和吸收波长的不匹配以及发射信号的重叠是荧光团(包括量子点)的物理特性，这是多重分析领域取得进展的关键障碍，无法消除。我们方法的一个关键特征是开发标记亲和产物，专为区分细胞表面和细胞内标记而设计，并针对电感耦合等离子体质谱检测。将开发可区分的稳定同位素元素标签的标记方法。该项目将提供稳定的金属标记、适当标记的亲和试剂、方法和用于区分多个分析物的示范数据。我们希望这种集成的试剂系统将极大地提高医生和他们的患者可获得的诊断、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经济成本。我们由公认的专家组成的跨学科小组将应对这一重大挑战。米切尔·A·温尼克教授和马克·尼茨教授将利用现有的能力来合成载体聚合物。将开发使用可区分的稳定同位素元素标签的方法。电感耦合等离子体质谱仪小组(弗拉基米尔·I·巴拉诺夫)将开发一种特定目的的分析方法，将机器人进样与电感耦合等离子体质谱仪器相结合，从而实现多重检测。Olga I.Ornatskaia将利用现有能力开发元素标记分析方法，并与常规分析方法进行不断的验证。

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.