Affinity Reagents and Sensor Platform Development for Blood Biochemical Monitoring

用于血液生化监测的亲和试剂和传感器平台开发

• 批准号: 10452054
• 负责人: Jing Pan
• 金额: $ 21.47万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452054
• 关键词: Address; Adopted; Affinity; Antibodies; Architecture; Binding; Biochemical; Biological Assay; Biological Markers; Biological Models; Biological Sciences; Biomedical Engineering; Biosensor; Blood; Blood Cells; COVID-19 patient; COVID-19 treatment; Caring; Cessation of life; Chronic; Chronic Disease; Clinical; Clinical Research; Cohort Studies; Conflict (Psychology); Critical Care; Critical Illness; Data; Databases; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Disease Management; Disease Progression; Dissociation; Drug Kinetics; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemiology; Equilibrium; Event; Fluorescence; Future; Goals; Healthcare; Healthcare Systems; Heating; Hospitals; Hour; Human; Immune; Immune System Diseases; Immune response; Immunoassay; Immunologics; Immunomodulators; Immunophenotyping; Immunotherapy; In Vitro; Individual; Inflammatory Response; Inpatients; Interleukin-6; Intervention; Knowledge; Libraries; Measurement; Medical; Microfluidic Microchips; Mission; Molecular; Monitor; Morbidity - disease rate; National Institute of Biomedical Imaging and Bioengineering; Oligonucleotides; Outcome; Patient Care; Patients; Performance; Phenotype; Physicians; Procedures; Process; Property; Proteins; Public Health; Reagent; Recycling; Research; Sampling; Scaffolding Protein; Scheme; Science; Scientist; Sepsis; Septic Shock; Signal Transduction; Surface; Survivors; Technology; Temperature; Therapeutic; Thrombin; Thrombomodulin; Time; Transducers; United States; Whole Blood; Work; Yeasts; absorption; aptamer; base; biomarker panel; care costs; clinical decision-making; clinical predictors; cytokine release syndrome; design; detection limit; diagnostic value; evidence base; experience; high risk; improved; individual patient; innovation; mortality; multiplex detection; nanoparticle; novel; novel therapeutics; operation; plasmonics; predictive tools; programmed cell death ligand 1; rate of change; real time monitoring; response; response to injury; scaffold; sensor; septic patients; severe COVID-19; technology development; tissue injury; treatment response

Project Summary/Abstract Sepsis is one of the major healthcare problems in the United States with an annual patient care cost approaching $23 billion. Despite a substantial decrease in inpatient mortality over the past 15 years, the current epidemiology indicates a disturbing increase in the number of sepsis survivors who develop a chronic critical illness (CCI) phenotype featuring prolonged ICU stay and poor long-term outcomes. The most important clinical challenge in sepsis care today is identifying early and continuously monitor those patients with a high risk of either dying or developing CCI. Previous clinical studies show unstable sepsis patients have distinct immunologic subtypes (endotype) defined by the level and fluctuation of circulating immunomodulators over time. Unfortunately, current standard immunoassays (e.g. ELISA) provide only intermittent sampling of biomarker levels with a sample-to-result time of several hours to days, which is insufficient for timely clinical decision making using immune endotypes. The long-term goal of this research is to develop improved bioanalytical tools for predicting clinical trajectory and tailoring medical intervention based on sepsis patient’s individual pathophysiologic endotype. The overall objective in this application is to develop a biosensor platform that addresses the analytical needs of immunologic endotyping in sepsis patient care. A major technical challenge in the real-time monitoring of biomolecules is achieving fast response while maintaining low limit of detection (LOD). To resolve this conflict, the application proposes to adopt the thermal cycling process used in PCR and repurpose it for real-time protein detection. We will develop affinity reagents with temperature-selective binding properties (aim 1). This novel affinity reagent will be integrated with an ultrafast photothermal cycling sensor platform to enable sensitive and frequent biomolecular monitoring (aim 2). In aim 1 we will use in vitro selection to identify binders with temperature-sensitive affinity towards a 3- biomarker panel containing two immune-modulating proteins (i.e. IL-6 and sPD-L1) and a tissue injury indicator thrombomodulin. The optimum temperature range and binding curve for the selected reagents will be characterized. In aim 2 we will develop a sensor platform capable of frequent, multiplexed detection of IL-6, sPD-L1 and thrombomodulin in human whole blood. The research proposed is innovative because it combines novel affinity reagents and sensing mechanisms to reconcile the slow off-rate needed for sensitive detection and the fast off-rate needed for fast assay time. The proposed technology is significant because it is generally applicable to many diseases associated with uncontrolled, systematic inflammatory response, such as the management of severe COVID-19 patients and treatment for patients with cytokine release syndrome after immunotherapy. Furthermore, the successful completion of this project will form the basis for future studies, such as the clinical study on immunologic endotypes diagnostic values at different stages of sepsis, the pharmacokinetics of novel therapeutics, and the closed-loop intervention of chronic diseases.

项目概要/摘要 败血症是美国主要的医疗保健问题之一，每年患者护理费用接近 23 美元 十亿。尽管过去 15 年住院患者死亡率大幅下降，但当前的流行病学表明 出现慢性危重病 (CCI) 表型的脓毒症幸存者数量令人不安地增加 ICU 住院时间延长，长期结果不佳。当今脓毒症护理中最重要的临床挑战是​​确定 尽早并持续监测那些死亡或发生 CCI 风险较高的患者。之前的临床研究 显示不稳定脓毒症患者具有不同的免疫亚型（内型），由免疫水平和波动定义 随着时间的推移循环免疫调节剂。不幸的是，当前的标准免疫测定（例如 ELISA）仅提供 生物标志物水平的间歇性采样，采样到结果的时间为数小时至数天，这对于 使用免疫内型及时做出临床决策。本研究的长期目标是开发改进的 用于预测临床轨迹并根据脓毒症患者个体定制医疗干预的生物分析工具 病理生理学内型。该应用的总体目标是开发一个生物传感器平台来解决 脓毒症患者护理中免疫内分型的分析需求。实时监控的一大技术挑战 生物分子正在实现快速响应，同时保持低检测限 (LOD)。为了解决这一冲突， 该申请建议采用 PCR 中使用的热循环过程，并将其重新用于实时蛋白质检测。我们 将开发具有温度选择性结合特性的亲和试剂（目标 1）。这种新型亲和试剂将 与超快光热循环传感器平台集成，可实现灵敏且频繁的生物分子监测 （目标 2）。在目标 1 中，我们将使用体外选择来识别对 3-生物标志物具有温度敏感亲和力的结合剂 包含两种免疫调节蛋白（即 IL-6 和 sPD-L1）和组织损伤指标血栓调节蛋白的组合。 将表征所选试剂的最佳温度范围和结合曲线。在目标 2 中，我们将开发 一个能够频繁、多重检测人全血中 IL-6、sPD-L1 和血栓调节蛋白的传感器平台。 所提出的研究具有创新性，因为它结合了新型亲和试剂和传感机制来协调 灵敏检测所需的慢解离速率和快速测定时间所需的快速解离速率。所提出的技术是 意义重大，因为它通常适用于许多与不受控制的系统性炎症相关的疾病 应对措施，例如重症 COVID-19 患者的管理和细胞因子释放综合征患者的治疗 免疫治疗后。此外，该项目的成功完成将为未来的研究奠定基础，例如 脓毒症不同阶段免疫内型诊断价值的临床研究、新型药物的药代动力学 治疗，以及慢性病的闭环干预。