A Cortisol Sensing Enzyme System: A New Platform Utilizing Dehydrogenases in Biosensors

项目摘要 第一阶段SBIR的目标是开发一种新的平台，使脱氢酶能够用作酶 在安培生物传感器中。无论平台或测量方式如何，所有生物传感器设计都需要 使用一种酶来正常发挥作用。生物传感器是测量化合物的设备，这些化合物对 人类健康。FDA已经批准了安培生物传感器，这种传感器可以监测人体内的血糖 目前正在开发几个新的生物传感器平台。生物传感器需要一种酶 这是要测量的化合物所特有的。 这项提议将通过开发一种酶系统来展示我们平台的效用，该系统能够提供 监测护理设备和生物传感器中的皮质醇。阻碍发展的主要障碍是 连续皮质醇监测系统是缺乏一种酶，可以用于设备制造的一部分。 目前，还没有描述适合于皮质醇检测和生物传感应用的酶 或者商业化。这说明了开发专门用于分析物的新的酶系统的重要性 对人类健康很重要。一旦在第一阶段演示了概念验证，我们的第二阶段将扩展 与其他脱氢酶一起使用的平台，用于监测对人类健康和疾病至关重要的分析物。阶段 II靶标包括马来酸脱氢酶、GABA脱氢酶、琥珀酸脱氢酶、丙酮酸 脱氢酶、乙醛脱氢酶和酒精脱氢酶。这些脱氢酶 要么没有已知的氧化酶对应物，要么有非常不稳定的氧化酶对应物，从而使 在研究自由活动的动物的生物传感器制造中，酶基本上毫无用处。在阶段结束时 II SBIR，我们将拥有一个优化的平台，生产高度稳定的脱氢酶，用于 生物传感器。此外，还将开发一种适用于人类的功能性皮质醇生物传感器。不是 目前存在连续测量体内皮质醇水平的诊断设备。 在这项研究中，我们将使用我们提出的平台来创建一个皮质醇酶系统，将两个新的 酶产生的信号可以在高灵敏度的电极上检测到。这种双酶 这种方法以共价连接的辅酶分子为特征，具有很高的创新性，将使 第一次，持续监测皮质醇等对人类健康重要的分析物。基于新设备的 这项技术将使临床医生能够监测和管理疼痛、压力和其他消极的情况 影响人类健康。我们的平台将对其他设备的开发和制造产生积极影响 用于新的传感应用。第一阶段的可交付物将是偶联酶系统的原型 演示了选择性、灵敏和高效的皮质醇水平监测机制的概念验证 组织和体液。这项提案的成功完成是建立 一种持续监测所有高危个体皮质醇的方法。

Project Abstract The goal of this Phase I SBIR is to develop a new platform that enables dehydrogenases to be used as enzymes in amperometric biosensors. Regardless of the platform or measurement modality, all biosensor designs require the use of an enzyme to properly function. Biosensors are devices that measure compounds important for human health. FDA approval has already been achieved for amperometric biosensors that monitor glucose in humans, and several new biosensor platforms are currently under development. Biosensors require an enzyme that is specific for the compound to be measured. This proposal will showcase our platform’s utility by developing an enzyme system that provides the ability to monitor cortisol in point-of-care devices and biosensors. The major impediment to the development of a continuous cortisol monitoring system is the lack of an enzyme that can be used as part of device fabrication. At this time, no enzyme suitable for cortisol detection and suitable for biosensing applications has been described or commercialized. This illustrates the importance of developing new enzyme systems specific for analytes important in human health. Once proof-of-concept is demonstrated in Phase I, our Phase II will extend the platform for use with other dehydrogenases to monitor analytes important in human health and disease. Phase II targets include maleate dehydrogenase, GABA dehydrogenase, succinate dehydrogenase, pyruvate dehydrogenase, acetaldehyde dehydrogenase, and alcohol dehydrogenase. These dehydrogenase enzymes either have no known oxidase counterpart or have oxidase counterparts that are so unstable as to render the enzyme essentially useless in biosensor fabrication for studies in freely moving animals. At the end of the Phase II SBIR, we will have an optimized platform that produces highly stabilized dehydrogenase enzymes for use in biosensors. In addition, a functioning cortisol biosensor suitable for use in humans will be developed. No diagnostic device presently exists to continuously measure cortisol level in vivo. In this study we will use our proposed platform to create a cortisol enzyme system by coupling together two novel enzymes to produce a signal that can be detected on an electrode with high sensitivity. This dual enzyme approach that features a covalently attached co-enzyme molecule, is highly innovative and will enable, for the first time, the continuous monitoring analytes important to human health such as cortisol. New devices based on this technology will allow clinicians to monitor and manage pain, stress, and other conditions that negatively impact human health. Our platform will positively impact the development and manufacturing of other devices for novel sensing applications. The Phase I deliverable will be a prototype of the coupled enzyme system demonstrating proof-of-concept for a selective, sensitive and efficient mechanism of monitoring cortisol levels in tissues and fluids. The successful completion of this proposal is the necessary first step to establishing a continuous method of monitoring cortisol in all at risk individuals.