Cross-species serum antibody detection by direct immunoglobulin catalysis

目前的医学实践中，通过观察症状结合血液检测来诊断莱姆病产生了很多错误。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。这项拟议的研究利用了一种对莱姆感染做出反应而形成的抗体的一种鲜为人知的特性，这种特性可以通过它们的化学反应能力方便地测量出来。莱姆病是美国最常见的媒介传播疾病，每年感染30万美国人。目前的医学实践是通过观察症状并结合用酶联免疫吸附试验和免疫印迹检测宿主动物的疏螺旋体抗体来诊断莱姆病。这两种方法都有其局限性，即二次试剂呈现非特异性结合。二次抗体使检测对提供血清的物种具有特异性。廉价、同型和物种无关的抗体传感器很重要，因为它们可以促进对传染病的广泛监测、血清转换动力学的跨物种研究和暴露后的早期检测。其长期目标是实现和标准化一种新的生物传感器，该传感器基于通过抗体催化的水氧化途径(ACWOP)直接测量抗体的催化作用。其目的是阐明抗原固定和催化传感产生过氧化氢用于比色或电化学读数的物理化学。将追求三个研究目标：即通过实验展示抗原定位和特定抗体在纸质基质上的结合；通过直接催化读出结合抗体来展示ELISA水平的敏感性；以及展示在临床相关样本中检测疏螺旋体抗原。关键特征是新的ACWOP转导机制，它消除了免疫特异性传感器中的二次抗体，并实际实现了抵抗非特异性结合的固定化化学。预计拟议的生物传感器将产生重大的实际影响，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宿主以及牲畜、伴生动物和人类。

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.