Multi-Channel Sensing using Fluorescent Protein-Nanoparticle Sensors

描述（由申请人提供）：“化学鼻/舌”方法提供了特异性识别和分离技术的潜在替代方案。在这种方法中，生成传感器阵列以通过选择性受体提供与分析物的差异相互作用，生成刺激响应模式，该刺激响应模式可以被统计分析并用于识别单个目标分析物以及复杂混合物的分析。最近，我们开发了纳米颗粒荧光聚合物传感器，用于通过荧光团置换机制识别蛋白质、细菌和癌细胞，以及用于生物流体中蛋白质检测的高灵敏度基于纳米颗粒GFP的“化学鼻”策略。 目前用于蛋白质和细胞表面的化学鼻传感器是单通道的，这意味着每个传感元件需要单独的孔或通道。这种对空间上不同的传感器元件的要求使得基于微板的技术和基于阵列的感测在其他场所（包括微流体平台和组织染色）中的应用变得复杂。为了克服这一限制，我们将利用荧光蛋白（FP）的光谱范围来提供传感应用的多通道荧光转导。多通道感测将促进基于阵列的感测的实现，允许“一个井”感测，同时通过比率计dadt的生成来提高灵敏度。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。 公共卫生相关性：蛋白质和细胞表面的快速有效传感将为许多疾病提供新的诊断方法。

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. PUBLIC HEALTH RELEVANCE: Rapid and efficient sensing of proteins and cell surfaces will provide access to new diagnostics for many diseases.