SBIR Phase II: Novel Microarray Platforms For Detection Of Rare Molecules In Complex Mixtures

这个小企业创新研究（SBIR）二期项目将为蛋白质组学应用提供优化的复合聚合物蛋白质结合表面。新的表面将专门设计用于反相蛋白质微阵列，以检测复杂生物混合物中的稀有分子。发现和量化复杂混合物中的稀有分子对于提高对疾病机制和进展以及对治疗方案的反应的理解至关重要。由于光学干扰、低蛋白质结合和非特异性相互作用的积累，目前在这些应用中使用的表面具有检测灵敏度有限的特性。本项目将优化并介绍一种用于蛋白质阵列应用的新型轨道蚀刻硝化纤维素复合膜的应用。将开发用于新复合材料的制造工艺，以产生多种形式的复合材料，使其能够并入各种结合分析格式。这项工作也将阐明产生超灵敏结合表面的重要性质。该项目的结果将是一种优化的复合膜，其特性和可制造性适合最敏感的结合应用，如逆相蛋白质阵列。该平台最初将优化用于复杂细胞裂解物中稀有分子的荧光检测。这个项目更广泛的影响/商业潜力将是提供一系列发现和诊断工具，扩大对人类疾病的了解、检测和治疗。目前临床试验中转化医学治疗的重点是确定个体患者中蛋白质（生物标志物）的表达模式。这些测量可以监测和了解个体化疾病的进展和对治疗的反应。它们将为制定有针对性的个性化治疗方案提供必要的数据。在过去的十年中，蛋白质阵列作为提供蛋白质表达的多重检测和定量的研究工具已经发现了实用性。然而，由于结合能力不足，动态范围有限，灵敏度差，这些工具作为提供患者特异性信息和指导药物治疗的诊断平台的全部潜力尚未实现。该项目定义了一种新的复合表面，当结合到多路免疫分析系统中时，它的结合能力和灵敏度都有显著的提高。该组合物可以包含在各种平台中，以增强个体尺度上重要标记物的发现和定量，以及用于广泛诊断应用的高通量系统。

This Small Business Innovation Research (SBIR) Phase II project will provide an optimized composite polymer protein binding surface for proteomics applications. The new surface will be specifically designed for reverse phase protein microarrays to enable detection of rare molecules in complex biological mixtures. Discovery and quantification of rare molecules in complex mixtures is essential to improve the understanding of disease mechanism and progression, and responses to treatment regimes. Current surfaces used in these applications have properties that exhibit limited sensitivity of detection due to optical interferences, low protein binding and accumulation of nonspecific interactions. This project will optimize and introduce the application of a new track etched , nitrocellulose composite membrane for protein array applications. Manufacturing processes for the new composite will be developed to generate multiple forms of the composite to allow it to be incorporated into a variety of binding assay formats. This effort will also shed light on important properties for generating ultrasensitive binding surfaces. The result of this project will be an optimized composite membrane with characteristics and manufacturability suited for the most sensitive binding applications, such as reverse phase protein arrays. The platform initially will be optimized for fluorescent detection of rare molecules in complex cell lysates. The broader impact/commercial potential of this project will be to provide a family of discovery and diagnostic tools that will expand the understanding, detection and treatment of human disease. The current focus in translational medicine for therapies in clinical trials is to identify expression patterns of proteins (biomarkers) in individual patients. These measurements allow the monitoring and understanding of individualized disease progression and responses to treatment. They will provide the data necessary to create targeted, personalized treatment regimens. Protein arrays have found utility over the past decade as research tools that provide multiplexed detection and quantification of protein expression. However, the full potential of these tools as diagnostic platforms that provide patient-specific information and guide drug treatment has not been realized due to insufficient binding capacity, limited dynamic range and poor sensitivity. This project defines a new composite surface that has a significant increase in both binding capacity and sensitivity when incorporated into multiplexed immunoassay systems. The composite can be included in a variety of platforms to enhance discovery and quantification of important markers on an individual scale as well as high throughput systems for broad diagnostic application.