Synthetic biological systems for protein detection

项目摘要 拟议工作的目标是创建一个新的基于合成生物学的蛋白质平台 诊断。激发这一挑战的特定背景是需要测量蛋白质 以最小设备的方式指示微量营养素缺乏的生物标记。当前 由于成本的原因，营养缺乏的诊断在所需的人口规模内是不可行的 和后勤限制，因此是一种护理点、最少设备、可现场部署的方法 需要帮助营养流行病学家获得他们需要的信息，以便更好地分配有限的 干预资源。然而，这种技术的影响将远远超出营养方面。 流行病学和诊断学，因为蛋白质是许多其他疾病和 条件。因此，同样的技术可以转化为全球健康应用程序，并 在资源匮乏的地区筛查其他疾病。此外，它还可以更好地实现 克服日常生活限制对新生儿重症监护病房患者的诊断监测 允许的抽血量。 分离蛋白报告系统与无细胞蛋白表达系统的使用是 提出要实现这一目标。在特定蛋白质存在的情况下，分离式报告系统 重新组装，反应发生色度变化。整个方法需要 最小甚至不需要设备，而且价格低廉，非常适合在低海拔地区使用- 受营养缺乏影响最大的资源区。有很强的 支持这种方法成功的可能性的初步数据。为了实现这些目标， 提出了三个目标。首先，将现有的传感器原理证明与模型相结合 蛋白质靶标和传感器设计选择对传感器功能和性能的影响 将被描述为。第二个目标需要通过以下方式来证明该方法的普适性 开发至少三种临床相关蛋白质传感器。第三个目标涉及推进 通过在人血清中测试其功能来实现真正的现场部署状态的测试 矩阵，实施校准方法，演示冷冻干燥后的功能，以及 开发一款配套的智能手机应用程序，以支持输出解释。 该项目将产生可用于有史以来第一个合成的基础技术 基于生物学的定量蛋白质检测分析，适用于资源匮乏的环境。通过低成本， 本质上是看护点，并且很容易推广到其他蛋白质靶点，这样一个长期结果 可能会改善全球数百万人的健康。

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.