Development of Sensitive Electrochemiluminescent Immunoassays through Electric Field Assisted Rapid Analyte Capture and Rational Design of the Reporter Catalyst/Reaction System

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This project is jointly funded by the Chemical Measurement and Imaging Program of the Division of Chemistry and the Established Program to Stimulate Competitive Research (EPSCoR) Program. Immunoassays offer a powerful approach to quantitating a variety of biological substances such as proteins, peptides, antibodies, and hormones, etc., in complex mixtures such as blood, saliva and sweat. Immunoassays are based on antigen-antibody interactions and as such, are very specific, capable of measuring very low concentrations. Professors Debashis Dutta and Michael Taylor of the University of Wyoming (UW) are developing electrochemiluminescent (ECL) immunoassays. The limits of measurement with this technique are over 1000-10,000 times lower than those using the current Enzyme Linked ImmunoSorbent Assays (ELISA). The development of even more rapid processes and new antigen-antibody interactions are central to this research. Education efforts include constructing a portable ECL instrument for use in the undergraduate classroom as well as developing a laboratory module for the undergraduate chemistry curriculum at UW. The team also continues to develop an outreach program with the Central Wyoming College (CWC). CWC enrolls significant numbers of Native Americans and first generation college students. The outreach activities with CWC include presentations on micro-/nanoscale analytical devices to the Science, Math and Engineering Club at CWC, as well as intensive on-site demonstrations of nanotechnology for the interested club member. These activities encourage the students to pursue careers in biotech and biomedical fields. This project focuses on the capture of antigen molecules on an electrode surface using a steady or a pulsating electric field. This technique improves the analyte capture efficiency as well as reduces the sample incubation period in the immunoassay. Moreover, the integration of an ECL detection method to the assay is pursued by triggering ECL reactions at the same electrodes. As part of this effort, the signal-to-noise ratio in the experiments may be improved through the synthesis of peptide-based catalyst oligomers which are chemically appended to well-defined locations on an antibody using cysteine-based conjugation chemistry. This project advances the national health and increases the participation of a wide range of students, including Native Americans.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.