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2-D Diffusion Assay on Polymer Brush for POC Cardiac Infarction Diagnosis

用于 POC 心肌梗死诊断的聚合物刷二维扩散测定

基本信息

批准号：
8512782
负责人：
Ashutosh Chilkoti
金额：
$ 18.21万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-17 至 2015-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The objective of this R21 proposal is to develop a multianalyte 2-D diffusion point-of-care assay (POC) for cardiac infarction. The 2-D diffusion assay consists of a "nonfouling" poly(oligoethyene glycol methacrylate (POEGMA) brush coated glass chip that contains two types of printed spots: "stable" spots of capture antibodies (Abc) and "soluble" spots of detection reagents that are printed with excipients so that they dissolve upon contact with blood. A finger stick of blood is applied to the surface, and the protein analytes in the blood diffuse across the nonfouling POEGMA brush and bind to immobile spots of the individual capture antibodies (Abc's) on the surface. Simultaneously, the blood dissolves the fluorescently labeled detection Ab's (Abd) from the labile spots, which then bind to their respective analyte-Abc spots, completing the sandwich. A final displacement step with buffer removes the blood components from the chip, revealing the formation of a fluorescent spot at the position where the unlabeled detection agent had been printed. This assay will require no sample preprocessing, will be multiplexed to detect cardiac troponin (cTnI) and the brain natriuretic pro-polypeptide (pro-BNP) on the same chip, and will provide quantitative read-out of analyte concentration, thereby providing a new dimension of capabilities to POC tests that are superior to the current gold standard - lateral-flow assays.

描述（由申请方提供）：本R21提案的目的是开发一种用于心肌梗死的多分析物二维扩散床旁检测（POC）。2-D扩散试验由“无污染”聚（低聚乙二醇甲基丙烯酸酯（POEGMA）刷涂的玻璃芯片组成，该芯片含有两种类型的打印点：捕获抗体（Abc）的“稳定”点和检测试剂的“可溶”点，这些点打印有赋形剂，以便它们在与血液接触时溶解。将手指针刺的血液施加到表面上，血液中的蛋白质分析物扩散穿过无污染的POEGMA刷，并结合到表面上的单个捕获抗体（Abc）的固定点。同时，血液溶解来自不稳定斑点的荧光标记的检测Ab（Abd），其然后结合到其各自的分析物-Abc斑点，完成夹心。用缓冲液进行的最终置换步骤从芯片上去除血液成分，揭示了在未标记的检测剂已被打印的位置处形成的荧光点。该检测试剂盒无需样本预处理，可在同一芯片上多重检测心肌肌钙蛋白（cTnI）和脑利钠肽原（pro-BNP），并可提供分析物浓度的定量读数，从而为POC检测提供上级当前金标准-侧流检测试剂盒的新功能。