Enzyme Amplified Array Sensing of Proteins for Cancer Detection

描述(由申请人提供)：用于癌症检测阵列检测的蛋白质的酶扩增阵列检测(“化学鼻/舌头”方法)为标准免疫检测策略提供了一种通用的替代方案。在这种方法中，产生一个传感器阵列，通过选择的受体提供与分析物的不同相互作用，产生刺激响应模式，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和复杂混合物。这种方法已经应用于生物传感，包括蛋白质和细胞表面。然而，由于这些传感器的灵敏度较低，其应用受到了限制。为了提供更高的灵敏度，我们将使用酶来创建基于阵列的传感器，以提供识别事件的放大。在这种酶扩增阵列传感(EAAS)方法中，阵列的灵敏度将通过酶反应来放大，将酶联免疫吸附试验的信号放大与“化学鼻子”方法的多功能性结合在一起。我们的研究将开发和优化EAAS传感器，并将它们应用于生物传感中的两个重要目标：目标1：我们将创建酶-纳米颗粒结合物来检测和识别蛋白质水平的变化，并根据裂解物中的蛋白质模式确定这些系统区分细胞类型和状态(例如，健康、癌症、转移)的能力。目的2：我们将使用小鼠模型来确定我们的传感器系统在利用组织样本检测和表型识别癌症方面的有效性。这项为期两年的R21拨款的目标是为蛋白质和细胞表面创造高灵敏度和高分辨的传感器，作为癌症诊断和病原体检测的平台。这些研究的结果将作为未来资助期动物和临床研究的基础。 公共卫生相关性：基于酶扩增阵列的传感为当前的免疫传感策略提供了一种通用和有效的替代方案。在我们提议的研究中，我们将开发直接适用于癌症诊断系统的蛋白质传感器。

DESCRIPTION (provided by applicant): Enzyme Amplified Array Sensing of Proteins for Cancer Detection Array-based sensing (the "chemical nose/tongue" approach) provides a versatile alternative to standard immunosensing strategies. 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 complex mixtures. This methodology has been applied to biosensing, including proteins and cell surfaces. Application of these sensors, however, has been limited due to their low sensitivity. To provide enhanced sensitivity, we will use enzymes to create array- based sensors that provide amplification of the recognition event. In this Enzyme Amplified Array Sensing (EAAS) approach, the sensitivity of the array will be amplified through an enzymatic reaction, coupling the signal amplification of ELISA with the versatility of the "chemical nose" approach. Our research will develop and optimize EAAS sensors and apply them to two important targets in biosensing: Aim 1: We will create enzyme-nanoparticle conjugates to detect and identify changes in protein levels, and determine the ability of these systems to differentiate cell types and states (e.g. healthy, cancerous, metastatic) based on protein patterns in lysate. Aim 2: We will use murine models to determine the effectiveness of our sensor system for detection and phenotypic identification of cancer using tissue samples. The goal of this two-year R21 grant is to create highly sensitive and discriminating sensors for proteins and cell surfaces that will serve as platforms for cancer diagnostics and pathogen detection. Results from these studies will be used as the foundation for animal and clinical studies in future funding periods. PUBLIC HEALTH RELEVANCE: Enzyme amplified array-based sensing provide a versatile and efficient alternative to current immunosensing strategies. In our proposed research we will develop sensors for proteins with direct applicability to diagnostic systems for cancer.