Cross-species serum antibody detection by direct immunoglobulin catalysis

直接免疫球蛋白催化跨物种血清抗体检测

• 批准号: 1706518
• 负责人: Brian Kirby
• 金额: $ 39.93万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1706518&HistoricalAwards=false
• 关键词: Cross; species; serum; antibody; detection

Current medical practice to diagnose Lyme disease by observation of symptoms combined with blood testing produces a lot of errors. The long-term goal is to develop a new approach that overcomes this limitation in Lyme disease testing and to create a general (cross­species) test for Lyme disease that can be used in humans, pets, livestock, and wild animals. The proposed research takes advantage of a largely-unknown property of the antibodies that form in response to Lyme infection that can be conveniently measured by their chemical reactivity. Lyme disease is the most common vector­borne disease in the United States, infecting 300,000 Americans per year. Current medical practice is to diagnose Lyme disease by observation of symptoms combined with serological detection of the host animal's antibodies against Borrelia antigens using ELISA and Western blot. Both of them have their limitations, namely, the secondary reagents exhibit nonspecific binding. The secondary antibodies make the test specific to the species providing the serum. Inexpensive, isotype- and species-independent antibody sensors are important because they can facilitate widespread monitoring of infectious disease, the cross-species study of the dynamics of seroconversion and early detection following exposure. The long-term goal is to implement and standardize a new biosensor based on direct measurement of antibody catalysis via the antibody-catalyzed water oxidation pathway (ACWOP). The objective is to elucidate the physiochemistry of antigen immobilization and catalytic sensing which produces hydrogen peroxide for a colorimetric or electrochemical readout. Three research aims will be pursued; namely, experimentally show antigen localization and specific antibody binding on paper substrates; show ELISA-level sensitivity from direct catalytic readout of bound antibodies; and, show detection of Borrelia antigens in clinically relevant samples. The key features are novel ACWOP-transduction mechanism that eliminates secondary antibodies from immune-specific sensors and practical implementation of immobilization chemistry that is resistant to nonspecific binding. It is anticipated that the proposed biosensor will have a significant practical impact because species­independent detection of Lyme borreliosis enables monitoring of the Borrelia reservoir as well as livestock, companion animals, and humans.

目前通过观察症状结合血液检测来诊断莱姆病的医疗实践产生了很多错误。 长期目标是开发一种新的方法，克服莱姆病测试中的这一限制，并创建一种可用于人类，宠物，牲畜和野生动物的莱姆病通用（跨物种）测试。这项拟议中的研究利用了抗体的一个很大程度上未知的特性，该特性是在对莱姆病感染做出反应时形成的，可以通过它们的化学反应性方便地测量。莱姆病是美国最常见的病媒传播疾病，每年感染30万美国人。 目前的医学实践是通过观察症状结合使用ELISA和Western印迹检测宿主动物针对疏螺旋体抗原的抗体来诊断莱姆病。它们都有其局限性，即第二试剂表现出非特异性结合。 第二抗体使测试对提供血清的物种具有特异性。廉价的、不依赖同种型和物种的抗体传感器很重要，因为它们可以促进传染病的广泛监测、血清转化动态的跨物种研究和接触后的早期检测。 长期目标是实现和标准化一种新的生物传感器，该生物传感器基于通过抗体催化水氧化途径（ACWOP）直接测量抗体催化。目的是阐明抗原固定和催化传感的生理化学，产生过氧化氢的比色或电化学读数。将追求三个研究目标;即，实验显示抗原定位和特异性抗体结合在纸基板上;显示ELISA水平的灵敏度从结合抗体的直接催化读出;和，显示在临床相关样品中的疏螺旋体抗原的检测。关键特征是新的ACWOP-转导机制，其从免疫特异性传感器中消除了二抗，并且实际实施了对非特异性结合具有抗性的固定化化学。预计所提出的生物传感器将具有显著的实际影响，因为莱姆疏螺旋体病的物种独立检测使得能够监测疏螺旋体储库以及牲畜、伴侣动物和人类。