Nanotube Reagentless Proteomic Arrays

描述(由申请人提供)：建议开发一种用于临床和生物样品中蛋白质浓度的无试剂多路测定的新技术。所提出的方法使用具有分子大小的开口的纳米管嵌入到机械和化学坚固的聚合物膜中。我们建议，可以使用这些膜构建微阵列，其中孤立的纳米管组被制成膜中的单独孔，每组/孔由不同的分子识别剂(MRA)衍生，例如抗体。在操作中，将首先在每个孔中发现具有对感兴趣蛋白质的MRA的阵列，然后将其暴露在包含一个或多个这些蛋白质分析物的分析物溶液中。混合物中的蛋白质分析物将与固定化的MRA形成结合伙伴(例如抗体/抗原复合体)。然后，阵列将通过一种新的显影和光学成像过程被“读出”，以产生与每个MRA相关的样本中的蛋白质浓度。为了与商业微阵列读取器兼容，阵列可以以显微镜载玻片的形式制成。在所提出的方法中，样品中的蛋白质被直接检测，不需要蛋白质分析物的预修饰或与标记的二次抗体形成夹心。此外，该方法具有单分子灵敏度(如果需要)。这项技术的核心概念已经由佛罗里达大学查尔斯·马丁教授的研究小组开发和演示。这项STTR工作将使马丁的团队与EIC实验室合作，为蛋白质组微阵列开发这项新技术。这项为期12个月的第一阶段研究旨在通过建立薄膜生产方法、演示潜在的光学成像现象、建立定量分析特征和变异系数以及演示光学成像格式来证明该方法的可行性。商业EIC实验室将通过制造和直接销售阵列作为毛坯和精选的MRA来实现技术商业化。市场和产品类似于现有的蛋白质微阵列产品，新方法提供了更快速和更灵敏的测试。建议的微阵列将在疾病检测/进展、蛋白质表达水平分析、小样本蛋白质分析(如活组织检查)、用于研究的蛋白质-蛋白质相互作用的鉴定、药物开发等领域、生物标记物发现等方面得到应用。这些应用已经指导了大量的蛋白质组微阵列市场；拟议的技术预计将在时间、成本和对现有方法的敏感性方面提供重大改进。

DESCRIPTION (provided by applicant): The development of a new technology for reagentless multiplexed determination of protein concentrations in clinical and biological samples is proposed. The proposed approach employs nanotubes with molecular sized openings embedded within a mechanical and chemically robust polymeric membrane. We propose that a microarray may be constructed using these membranes in which isolated groups of nanotubes are fabricated as separate wells in the membrane, each group/well derivatized with a different molecular recognition agent (MRA), such as an antibody. In operation, an array would be spotted first in each well with MRAs to the proteins of interest, and then exposed to the analyte solution containing one or more of these protein analytes. The protein analytes in the mixture would form binding partners (e.g., antibody/antigen complex) with the immobilized MRAs. The array would then be "read out" by a novel development and optical imaging process, to yield the protein concentration in the sample associated with each MRA. The arrays can be made in microscope slide format for compatibility with commercial microarray readers. In the proposed approach, proteins in a sample are detected directly, with no protein analyte premodification or sandwich formation with a labeled secondary antibody. Furthermore, the method is capable of single molecule sensitivity (if desired). The core concepts of this technology have already been developed and demonstrated by Prof. Charles Martin's research group at the University of Florida. This STTR effort will team Martin's group with EIC Laboratories to develop this new technology for proteomic microarrays. The 12 month Phase I study is designed to demonstrate the feasibility of the approach by establishing membrane production methods, demonstrating the underlying optical imaging phenomenon, establishing quantitative assay characteristics and coefficients of variation, and demonstrating optical imaging formats. Commercial EIC Laboratories will commercialize the technology by manufacturing and direct sales of the arrays as blanks and with selected MRAs. The market and products are similar to existing protein microarray products, with the new approach offering more rapid and sensitive testing. The proposed microarrays will find applications in disease detection/progression, analysis of protein expression levels, protein analysis for small samples such as biopsies, identification of protein-protein interactions for research, drug development, etc., biomarker discovery. These applications already guide the substantial proteomic microarray market; the proposed technology is expected to offer significant improvements in time, cost and sensitivity to existing approaches.