Developing a Small-Molecule Immunoassay

在化学系化学测量和成像计划的支持下，犹他大学的康博伊教授正在为小分子(低分子物质)的检测和定量开发新的分析方法。抗体具有化学特异性和高亲和力，特别适合于生物特异性的检测和定量。然而，利用抗体对低分子物质进行直接无标记检测是一个主要的技术挑战。康博伊教授正在使用非线性光学方法开发一种高度灵敏的小分子免疫分析方法，为检测和定量低分子物质提供一种可靠的、无标记的方法。研究成果是提供一套新的高度灵敏的分析工具，用于非法物质的检测和有害物质的筛选。在研究目标中整合了分析/生物分析化学的研究生和本科生的教育，这对保持美国经济的实力和美国生物技术产业的竞争优势具有重要的全国意义。康博伊教授还参与了演示、科学博览会活动，并协助在盐湖谷当地小学建立专门的科学实验教学空间，目的是让小学生接触到化学世界的奇观，促进教师培训和向年轻人介绍科学潜力。拟议研究的科学目标是实施二次谐波相关光谱(SHCS)用于快速单点校准，并利用二次谐波成像(SHI)的高通量能力进行筛选和量化。与现有方法相比，这些互补的技术、SHCS和SHI具有许多固有的优势。SHCS具有固有的优势，即可以从单一浓度的分析物获得校准(结合亲和力数据)，以在传统热力学平衡结合等温线测量所需时间的一小部分内得出Kd(或Ka)，并具有获得缔合和解离速率的额外好处。SHI具有利用抗体捕获阵列进行高通量测量的优势。这两种方法在生物学上也是通用的，能够检测任何包含共轭结构的分子，包括目前检测中几乎所有的生物分子。总而言之，这些方法代表了一种独特的分析方法，既不影响灵敏度和速度，又提供了一种新颖、经济高效和灵活的免疫分析格式。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Conboy at the University of Utah is developing new analytical methods for small-molecule (low molecular weight substances) detection and quantification. Antibodies, with their chemical specificity and high affinity are exceptionally suited for bio-specific detection and quantification. However, utilizing antibodies for the direct label-free detection of low molecular weight substances is a major technical challenge. Professor Conboy is using nonlinear optical methods to develop a highly sensitive small-molecule immunoassay, providing a reliable, label-free method for detecting and quantifying low molecular weight substances. The research outcome is to provide a set of new highly sensitive analytical tools for the purpose of detection of illicit substances and hazardous substances screening. Integrated within the research objectives is the education of both graduate and undergraduate students in analytical/bioanalytical chemistry, which is of great National importance in maintaining the strength of the US economy and the competitive edge of US biotechnology industries. Professor Conboy is also involved in hands on demonstrations, science fair activities and assistance with setting up dedicated teaching space for science experiments at local primary schools in the Salt Lake valley with the goal of exposing primary grade students to the wonders of the chemical world and to promote teacher training and introducing young people to the potentials of science.The scientific goal of the proposed studies is to implement second-harmonic correlation spectroscopy (SHCS) for rapid single point calibration and utilize the high-throughput capabilities of second harmonic imaging (SHI) for screening and quantification. These complimentary techniques, SHCS and SHI have many inherent advantages over existing approaches. SHCS has the intrinsic advantage that calibration (binding affinity data) can be obtained from a single concentration of the analyte to derive the Kd (or Ka) in a fraction of the time required by a conventional thermodynamic equilibrium binding isotherm measurement, with the added benefit of acquiring both the association and dissociation rates. SHI has the advantage of high-throughput measurement by utilizing antibody capture arrays. Both methods are also biologically versatile and capable of detecting any molecule containing conjugated structures, including virtually all biomolecules targeted in current assays. Together these methods represent a unique analytical approach that does not compromise sensitivity and speed yet provides a novel, cost effective, and flexible immunoassay format.