Multi-Channel Sensing using Fluorescent Protein-Nanoparticle Sensors

描述（由申请人提供）：“化学鼻/舌”方法提供了特定识别和分离技术的潜在替代方案。在这种方法中，产生传感器阵列，通过选择性受体提供与分析物的差异相互作用，产生刺激反应模式，该模式可以进行统计分析，并用于识别单个目标分析物和分析复杂混合物。最近，我们开发了纳米颗粒-荧光聚合物传感器，用于通过荧光团位移机制识别蛋白质，细菌和癌细胞，以及基于高灵敏度纳米颗粒-绿色荧光蛋白的“化学鼻子”策略，用于生物流体中的蛋白质检测。目前用于蛋白质和细胞表面的化学鼻子传感器是单通道的，这意味着每个传感元件都需要一个单独的孔或通道。这种对空间上不同的传感器元件的要求使基于微板的技术和基于阵列的传感在其他场所的应用复杂化，包括微流体平台和组织染色。为了克服这一限制，我们将利用荧光蛋白（FPs）的光谱范围为传感应用提供多通道荧光转导。多通道传感将促进阵列传感的实施，允许“一井”传感，同时通过生成比率测量数据提高灵敏度。在这个项目中，我们将使用超分子化学的工具来协同设计蛋白质和AuNP猝灭剂，为蛋白质和细胞表面传感提供高效和通用的平台。

DESCRIPTION (provided by applicant): The "chemical nose/tongue" approach presents a potential alternative to specific recognition and separations techniques. In this approach a sensor array is generated to provide differential interaction with analytes via selective receptors, generating a stimulus response pattern that can be statistically analyzed and used for the identification of individual target analytes and also analysis of complex mixtures. Recently, we have developed nanoparticle-fluorescent polymer sensors for identification of proteins, bacteria, and cancerous cells through a fluorophore-displacement mechanism as well as a highly sensitive nanoparticle-GFP based "chemical nose" strategy for protein detection in biofluid. Current chemical nose sensors for proteins and cell-surfaces are single channel, meaning a separate well or channel is required for each sensing element. This requirement for spatially distinct sensor elements complicates both microplate-based techniques and the application of array-based sensing in other venues, including microfluidics platforms and tissue staining. To overcome this limitation, we will exploit the spectral range of fluorescent proteins (FPs) to provide multi-channel fluorescence transduction for sensing applications. Multi-channel sensing will facilitate implementation of array-based sensing, allowing "one well" sensing while improving sensitivity through generation of ratiometric dadt. In this program, we will use the tools of supramolecular chemistry to synergistically engineer both the protein and AuNP quencher to provide highly efficient and versatile platforms for protein and cell surface sensing.