Protein-folding-based in-vivo biosensors

基于蛋白质折叠的体内生物传感器

基本信息

批准号：
10063408
负责人：
Kevin W Plaxco
金额：
$ 22.65万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10063408
关键词：
Adopted Affinity Animal Model Antibiotics Antibodies Binding Biological Markers Biosensor Blood Blood Vessels Brain Chemicals Clinical Corpus striatum structure Couples Cyclosporine DNA Detection Development Devices Diagnostic Disease Dose Drug Kinetics Electrodes Electron Transport Elements Engineering Feedback Foundations Frequencies Glucose Goals Health Hormones Hour Immunization Immunomodulators Immunosuppressive Agents In Situ In Vitro Inferior Interleukin-6 Measurable Measurement Measures Medical Research Membrane Proteins Metabolic Methotrexate Methylene blue Molecular Molecular Conformation Molecular Target Monitor Nucleic Acids Oligonucleotides Outcome Output Oxidation-Reduction Performance Phage Display Pharmaceutical Preparations Physiological Precision therapeutics Preclinical Testing Progressive Disease Proteins Rattus Reporter Reporting Research Research Project Grants Resolution Risk Sepsis Signal Transduction Specificity Techniques Technology Testing Therapeutic Index Time Uncertainty Validation Whole Blood Work aptamer awake base body system clinical practice cytokine design diagnostic biomarker glucose monitor improved in vivo in vivo evaluation innovation minimally invasive nanobodies new technology novel strategies oxidation precision drugs prevent programs protein biomarkers protein folding real time monitoring receptor response sensor sensor technology small molecule success temporal measurement

项目摘要

Summary. Our overarching research goal is the development of a minimally invasive, receptor-based technology able to monitor the concentration of effectively any specific molecule (irrespective of its chemical reactivity) in the living body. To this end, we have already demonstrated the ability of electrochemical, aptamer-based (E-AB) sensors to monitor a range of drugs and metabolites in situ in the blood vessels and brain of awake, freely moving rats with seconds resolution and measurement durations of hours. A potentially significant limitation of the platform, however, is its reliance on nucleic acid aptamers, the limited chemical complexity of which will no doubt ultimately restrict the number of targets amenable to detection using the approach. In response, we propose here to develop the first sensors in this class that instead employ proteins as their recognition elements. Specifically, we propose two aims that will significantly de-risk the development of protein-folding-based electrochemical sensors, setting the stage for their development as a powerful new approach to molecular monitoring. First, we will expand beyond the single, proof-of-principle example (a sensor fabricated using the FynSH3 domain as our receptor) we have realized to date to sensors against three additional, clinically important targets (the sepsis-diagnostic cytokine IL-6, the chemotherapeutic and immune modulator methotrexate, and the immunosuppressant cyclosporine) as evidence that our design strategy is general. Second, we will adapt these sensors, which we have already shown are capable of multi-hour performance in undiluted whole blood in vitro, to the more challenging measurement conditions found in vivo. If successful, the R21-scale project described here will lay the foundation for an R01-level research program that couples this technology with in vitro and in vivo protein selection to create sensors supporting the continuous, real time measurement of many clinically important molecules, including narrow-therapeutic-index drugs and protein biomarkers indicative of the status of many grievous, rapidly progressive diseases.

概括。我们的总体研究目标是开发基于受体的微创，基于受体能够监测有效任何特定分子的浓度的技术（不论其化学物质如何反应性）在活体内。为此，我们已经证明了电化学的能力，基于适体的（E-AB）传感器，以监测血管中的一系列药物和代谢物，并清醒的大脑，自由移动大鼠，分辨率和测量时间的时间为几个小时。潜在的但是，该平台的显着限制是它依赖核酸适体，有限的化学物质毫无疑问，其复杂性最终将限制使用使用该目标的目标数量方法。作为回应，我们在这里建议开发此类中的第一个传感器，而不是采用蛋白质作为他们的认可元素。具体而言，我们提出的两个目标将大大降低发展的危险基于蛋白质折叠的电化学传感器，为它们的发展奠定了基础分子监测的方法。首先，我们将扩展超越单一的原则示例（传感器使用Fynsh3域作为我们的受体制造）我们已经意识到迄今为止，传感器针对三个附加临床上重要的靶标（败血症诊断细胞因子IL-6，化学治疗和免疫甲氨蝶呤调节剂和免疫抑制剂环孢素）作为我们的设计策略的证据一般的。其次，我们将适应这些传感器，我们已经显示的能够多小时在体外未稀释的全血液中的表现，在体内发现的更具挑战性的测量条件。如果成功的R21级项目此处描述的项目将为R01级研究计划奠定基础将这项技术与体外和体内蛋白质选择相结合，以创建支持连续的，实时测量许多临床上重要的分子，包括狭窄的治疗性指数药物和蛋白质生物标志物指示许多严重的，快速进行性疾病的状态。