Troponin Biosensor for Early Detection and Real-time Monitoring of Myocardial Infarction

项目摘要 心血管疾病(CVD)是世界上大多数种族和民族的主要死因 美国。急性心肌梗死(AMI)(也称为心脏病发作)是心血管疾病的首要死亡原因。 早期发现急性心肌梗死可以转化为更早的干预，并提高存活率。当心肌细胞 受损的心肌肌钙蛋白(CTn)是心肌的一种成分，被释放到循环中。动态和 血cTn水平的递增升高预示着急性心肌梗死心脏损伤的持续事件 心肌细胞受损。因此，血液中的cTn水平通常用商业cTn免疫分析法来测量。 疑似患有急性心肌梗死的患者。然而，目前的cTn免疫分析方法在灵敏度和速度上存在局限性。 由于目前的cTn免疫分析中使用了两种抗体(一种作为捕获抗体，另一种作为检测抗体 试剂)，目前的cTn免疫分析需要多步反应，灵敏度有限。此外， 化验不是实时进行的，通常需要大约30分钟才能得到可能错过早期结果的结果 CTn水平或其变化率升高，从而延误诊断。EZ-Lab，与调查人员合作 奥克兰大学、韦恩州立大学和大学。密苏里州哥伦比亚大学的一位科学家建议开发一种新型的cTn生物传感器 基于我们多年的研究和开发，可实现高灵敏度(pg/ml)和实时检测cTn 1)独特设计的多肽模拟表位生物传感接口，用于无标记亲和力 2)小型化、低成本、实时的电化学传感器平台。在尖锐的 与目前使用两种抗体的cTn免疫分析相比，我们成功地证明了该多肽 当通过自组装单层(SAM)固定在表面时，模拟表位可以代替抗体 显著降低结构的可变性，保留生物活性，最大限度地减少或消除非特异性 从干扰蛋白质中吸附，并提供实时、高灵敏度和选择性的蛋白质检测 人类血清样本中的抗原。将这一新型多肽模拟表位生物传感技术推广到早期 检测和监测急性心肌梗死的临床应用，我们将合理设计多肽，形成强大的cTn 用于电化学cTn传感器的生物传感接口，可一步实时检测人体cTn 血清和血液样本的三个研究目标：1.开发高灵敏的多肽传感接口 用于cTn的检测；2.验证基于多肽SAM的cTn生物传感器的分析性能 血清和血液样本；3.血液样本中cTn的实时检测。建议的多肽模拟表位cTn 生物传感器有望具有高灵敏度和高定量，比目前用于 临床，允许以显著较低的成本早期诊断急性心肌梗死，在现实世界中量化cTn生物标记物 临床样本。我们的简单易用的cTn生物传感器有望用于监测患者的心脏 损伤状态，例如由紧急医疗服务团队从以下位置获取心脏损伤或急性心肌梗死的关键数据 帮助患者立即制定紧急救生治疗和管理计划。

Project Summary Cardiovascular disease (CVD) is the leading cause of death for people in most racial and ethnic groups in the United States. Acute Myocardial Infarction (AMI) (also called heart attack) is the top cause of death in CVD. Early detection of AMI translates to earlier intervention and improves survival rates. When cardiac myocytes are damaged, cardiac troponin (cTn), a component of the heart muscle, is released into circulation. Dynamic and incremental elevation of blood cTn levels is indicative of an on-going event of heart injury with acute cardiomyocyte damage. Thus, blood cTn level is routinely measured with commercial cTn immunoassays in patients suspected of having AMI. However, current cTn immunoassays are limited in sensitivity and speed. Since two antibodies are used in the current cTn immunoassays (one as the capture and another as the detection agents), current cTn immunoassays require multiple step reactions with limited sensitivity. Additionally, the assays are not performed in real time and typically take about 30 mins to get results that could miss the early rises in cTn levels or its rate of change thus delaying diagnosis. EZ-Lab, in collaborations with investigators at Oakland Univ., Wayne State Univ., and Univ. of Missouri-Columbia, proposes to develop a novel cTn biosensor that allows highly sensitive (pg/ml) and real-time detection of cTn based on our years of research and innovations in 1) uniquely designed peptide mimotope biosensing interface for label-free affinity based electrochemical biosensor; 2) miniaturized, low cost and real-time electrochemical sensor platform. In sharp contrast to current cTn immunoassays using two antibodies, we successfully demonstrated that peptide mimotopes, in lieu of antibodies, when immobilized on the surface via self-assemble monolayer (SAM) can significantly reduce the structural variability, retain biological activity, and minimize or eliminate non-specific adsorption from interfering proteins and provide real-time, highly sensitive and selective detection of protein antigens in human serum samples. To advance this novel peptide mimotope biosensing technology for early detection and monitoring of AMI for clinical applications, we will rationally design peptides to form robust cTn biosensing interface for electrochemical cTn sensors allowing one step and real-time detection of cTn in human serum and blood samples with three research Aims: 1. Develop a highly sensitive peptide sensing interface for detection of cTn; 2. Validate peptide SAM based cTn biosensor analytical performance using human serum and blood samples; 3. Real-time cTn sensing in blood sample. The proposed peptide mimotope cTn biosensor is expected to be highly sensitive and quantitative, faster than current cTn immunoassays used in the clinics, allowing for early diagnosis of AMI at significantly lower cost for quantifying cTn biomarkers in real world clinical samples. It is expected that our easy-to-use cTn biosensor can be used to monitor patients’ cardiac injury status such as by emergency medical service team to obtain the critical data on cardiac injury or AMI from the patients instantly to help develop urgent and life-saving treatment and management plan.