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

该项目由化学部的化学测量和成像计划和刺激竞争性研究的既定计划（EPSCoR）计划共同资助。免疫分析提供了一种强大的方法来定量各种生物物质，如蛋白质、多肽、抗体和激素等，在复杂的混合物，如血液、唾液和汗液。免疫测定是基于抗原-抗体相互作用，因此，是非常特异性的，能够测量非常低的浓度。怀俄明大学（UW）的Debashis Dutta和Michael Taylor教授正在开发电化学发光（ECL）免疫测定法。该技术的测量限比目前使用的酶联免疫吸附测定法（ELISA）低1000- 10000倍。发展更快速的过程和新的抗原-抗体相互作用是本研究的核心。教育方面的努力包括建造一个用于本科课堂的便携式ECL仪器，以及为西澳大学本科化学课程开发一个实验室模块。该团队还继续与中央怀俄明学院（CWC）开展外展计划。CWC招收了大量的美国原住民和第一代大学生。与CWC合作的外展活动包括向CWC的科学、数学和工程俱乐部介绍微/纳米级分析设备，以及为感兴趣的俱乐部成员提供密集的纳米技术现场演示。这些活动鼓励学生从事生物技术和生物医学领域的职业。这个项目的重点是利用稳定或脉动电场捕获电极表面的抗原分子。该技术提高了分析物捕获效率，并减少了免疫分析中的样品潜伏期。此外，通过在相同的电极上触发ECL反应，将ECL检测方法整合到分析中。作为这项工作的一部分，实验中的信噪比可以通过合成基于半胱氨酸的催化剂低聚物来提高，这些低聚物通过化学方法附加在抗体上明确的位置上。该项目促进了国民健康，并增加了包括印第安人在内的广大学生的参与。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.