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Multiplexed Ultrasensitive Mix-and-Read Tumor Marker Quantification Assays

多重超灵敏混合读取肿瘤标志物定量分析

基本信息

批准号：
7107550
负责人：
ARMIN H REITMAIR
金额：
$ 10万
依托单位：
NESHER TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-05-10 至 2008-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7107550
关键词：
antibody neoplasm /cancer proteins reading

项目摘要

DESCRIPTION (provided by applicant): Nesher Technologies, Inc. (NTI) licensed from UCLA the IP for a revolutionary suite of ultrasensitive biodetection technologies involving alternating laser excitation (ALEX) fluorescence spectroscopy. These technologies were developed at the UCLA Single Molecule Biophysics Laboratory (headed by Prof. Shimon Weiss). NTI proposes to apply one of these technologies for very early-stage cancer diagnosis, monitoring of therapeutic response, assessment of outcome, prediction of recurrence, as well as basic cancer research. In the proposed implementation, dubbed alternating-laser excitation fluorescence-aided molecule sorting, or ALEX-FAMS, two recognition molecules are tagged with different color fluorescence dyes. Coincident detection of the two colors in a femto-liter confocal detection volume constitutes a positive target detection event. The proposal's broad long-term objective is development of highly multiplexed, mix-and-read, high- throughput solution phase assays for rapid and ultra-sensitive (femtomolar) quantification of protein and mRNA levels of tumor markers in serum, urine, and saliva (with a capacity of >10,000 markers per standard patient sample), overcoming drawbacks of current technologies and minimizing costs. Over the funding period, we will develop a protein-based assay using two high affinity fluorescently labeled antibodies to detect carcino-embryonic antigen (CEA) and demonstrate the quantitative nature of our methodology. Our specific aims are: 1. Expression and purification of a high-affinity antibody pair specific for CEA 2. 2. Conjugation of two different fluorescent dyes to these antibodies 3. Evaluation of CEA detection with ALEX-based single molecule fluorescence spectroscopy Development of ultra-sensitive, low-cost diagnostic assays that radically push the limits of current technologies by allowing simultaneous detection and accurate quantification of thousands of tumor markers from a single patient sample will aid both clinicians and cancer researchers in improving usefulness of tumor marker analysis. The prospect of monitoring the public on a routine basis for cancer-related changes in bodily fluids will significantly decrease mortality rates and and help fighting this deadly disease.

描述（由申请人提供）：Nesher Technologies, Inc. (NTI) 已从加州大学洛杉矶分校 (UCLA) 获得知识产权许可，用于一套革命性的超灵敏生物检测技术，涉及交替激光激发 (ALEX) 荧光光谱。这些技术是在加州大学洛杉矶分校单分子生物物理实验室（由 Shimon Weiss 教授领导）开发的。 NTI 提议将其中一项技术应用于极早期癌症诊断、治疗反应监测、结果评估、复发预测以及基础癌症研究。在所提出的实施方案中，称为交替激光激发荧光辅助分子分选（ALEX-FAMS），两个识别分子用不同颜色的荧光染料标记。在飞升共焦检测体积中同时检测到两种颜色构成了阳性目标检测事件。该提案的广泛长期目标是开发高度多重、混合读取、高通量溶液相测定，用于快速、超灵敏（飞摩尔）定量血清、尿液和唾液中肿瘤标志物的蛋白质和 mRNA 水平（每个标准患者样本的容量超过 10,000 个标志物），克服当前技术的缺点并最大限度地降低成本。在资助期间，我们将开发一种基于蛋白质的检测方法，使用两种高亲和力荧光标记抗体来检测癌胚抗原（CEA），并证明我们方法的定量性质。我们的具体目标是： 1. CEA 2 特异性高亲和力抗体对的表达和纯化。 2. 两种不同荧光染料与这些抗体的缀合 3. 基于 ALEX 的单分子荧光光谱法检测 CEA 的评估开发超灵敏、低成本的诊断检测方法，通过同时检测和准确定量单个患者样本中的数千种肿瘤标志物，从根本上突破了当前技术的极限，将有助于临床医生和癌症研究人员提高肿瘤标志物分析的实用性。定期监测公众体液中与癌症相关的变化的前景将显着降低死亡率，并有助于对抗这种致命的疾病。