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

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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.