Synthetic biological systems for protein detection

用于蛋白质检测的合成生物系统

• 批准号: 10639348
• 负责人: Mark Philip-Walter Styczynski
• 金额: $ 35.6万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10639348
• 关键词: Affect; Antibodies; Base Sequence; Biological Assay; Biological Markers; Blood; Blood Tests; Blood Volume; C-reactive protein; Calibration; Cells; Cessation of life; Characteristics; Child; Chimeric Proteins; Clinical; Companions; Complex; Coupled; DNA-Directed RNA Polymerase; Data; Detection; Development; Diagnostic; Diagnostic Equipment; Dietary Intervention; Dietary intake; Disease; Electricity; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemiologist; Epidemiology; Equipment; Ferritin; Freeze Drying; Friends; Future; Goals; Growth; Health; Hour; Human; Hydration status; Impairment; Infant; Inflammation; Intervention; Laboratories; Letters; Literature; Location; Malnutrition; Mass Spectrum Analysis; Measurement; Measures; Medical; Micronutrients; Minerals; Modeling; Monitor; Neonatal Intensive Care Units; Nutrient; Nutritional; Nutritional status; Output; Patients; Performance; Persons; Policy Making; Population; Populations at Risk; Proteins; Reaction; Reporter; Resource-limited setting; Resources; Sampling; Serum; Site; System; Techniques; Technology; Testing; Time; Translating; Visual; Vitamins; Work; Zinc; biological systems; burden of illness; clinical biomarkers; clinically relevant; cost; design; detection assay; diagnostic value; epidemiology study; frontier; global health; health application; improved; insight; iron deficiency; lateral flow assay; micronutrient deficiency; novel strategies; nutritional epidemiology; point of care; portability; protein biomarkers; protein expression; screening; sensor; small molecule; smartphone application; success; synthetic biology

Project Summary The goal of the proposed work is to create a new synthetic biology-based platform for protein diagnostics. The specific context motivating this challenge is the need to measure protein biomarkers indicative of micronutrient deficiencies in a minimal-equipment fashion. Current diagnostics for nutritional deficiencies are infeasible at the required population scale due to cost and logistical constraints, and so a point-of-care, minimal-equipment, field-deployable approach is needed to help nutritional epidemiologists get the information they need to better allocate limited intervention resources. However, the impact of such a technology would go well beyond nutritional epidemiology and diagnostics, as proteins are common biomarkers for many other diseases and conditions. Thus, the same technology could be translated to global health applications and screening for other diseases in resource-poor locations. In addition, it could also enable better diagnostic monitoring for neonatal intensive care unit patients by overcoming limitations on daily allowable blood draw volumes. The use of a split protein reporter system coupled with a cell-free protein expression system is proposed to accomplish this goal. In the presence of a specific protein, the split reporter system reassembles and the reaction undergoes a colorimetric change. The entire approach requires minimal to no equipment and would be inexpensive, making it perfectly suited for use in the low- resource regions that are most prominently affected by nutritional deficiencies. There is strong preliminary data supporting the likelihood of success for this approach. To achieve these goals, three aims are proposed. First, the existing proof of principle sensor will be advanced with a model protein target and the impact of sensor design choices on sensor functionality and performance will be characterized. The second aim entails demonstrating the approach’s generalizability by developing at least three sensors for clinically relevant proteins. The third aim involves advancing the assay towards a truly field-deployable state by testing it for functionality in a human serum matrix, implementing a calibration approach, demonstrating functionality after lyophilization, and developing a companion smartphone app to support output interpretation. This project will yield the underlying technology that can be used for the first-ever synthetic biology-based, quantitative protein detection assay for low-resource settings. By being low-cost, essentially point-of-care, and easily generalizable to other protein targets, such a long-term result would potentially improve the health of millions of people worldwide.

项目摘要 这项工作的目标是建立一个新的基于合成生物学的蛋白质平台， 诊断激发这一挑战的具体背景是需要测量蛋白质 以最少的设备方式提供指示微量营养素缺乏的生物标志物。电流 由于成本问题，在所需的人口规模上，营养缺乏症的诊断是不可行的。 和后勤限制，因此，一个即时护理，最低设备，现场部署的方法 需要帮助营养流行病学家获得他们需要的信息，以更好地分配有限的 干预资源。然而，这种技术的影响将远远超出营养范围 流行病学和诊断，因为蛋白质是许多其他疾病的常见生物标志物， 条件因此，同样的技术可以应用于全球健康， 在资源贫乏地区筛查其他疾病。此外，它还可以更好地 通过克服日常检查的局限性对新生儿重症监护病房患者进行诊断监测 允许的采血量。 使用与无细胞蛋白表达系统偶联的裂解蛋白报告系统， 建议实现这一目标。在特定蛋白质的存在下，分裂报告系统 重新组装，反应发生比色变化。整个方法需要 最小到没有设备，而且价格便宜，非常适合在低海拔地区使用。 受营养不良影响最严重的资源区。有强有力 支持这种方法成功可能性的初步数据。为了实现这些目标， 提出了三个目标。首先，现有的原理传感器的证明将与模型先进 蛋白质靶标以及传感器设计选择对传感器功能和性能的影响 将被定性。第二个目标需要通过以下方式证明该方法的普遍性： 为临床相关蛋白质开发至少三种传感器。第三个目标是推进 通过测试其在人血清中的功能性， 基质，实施校准方法，证明冻干后的功能性，以及 开发配套的智能手机应用程序来支持输出解释。 该项目将产生可用于有史以来第一次合成的基础技术 基于生物学的定量蛋白质检测分析，适用于低资源环境。通过低成本， 基本上是即时的，并且容易推广到其他蛋白质靶点，这样的长期结果 将有可能改善全世界数百万人的健康状况。