Nanotube Reagentless Proteomic Arrays

描述（由申请人提供）：拟定开发一种新技术，用于临床和生物样本中蛋白质浓度的无试剂多重测定。所提出的方法采用嵌入在机械和化学上坚固的聚合物膜内的具有分子大小的开口的纳米管。我们提出，可以使用这些膜来构建微阵列，其中分离的纳米管组被制造为膜中的单独的威尔斯孔，每个组/孔用不同的分子识别剂（MRA）如抗体衍生化。在操作中，首先将阵列点样在具有针对感兴趣蛋白质的MRA的每个孔中，然后暴露于含有这些蛋白质分析物中的一种或多种的分析物溶液。混合物中的蛋白质分析物将形成结合配偶体（例如，抗体/抗原复合物）与固定的MRA。然后，通过一种新的显影和光学成像过程来“读出”阵列，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形式制备。在所提出的方法中，直接检测样品中的蛋白质，没有蛋白质分析物预修饰或与标记的二抗形成夹心。此外，该方法能够具有单分子灵敏度（如果需要）。这项技术的核心概念已经由佛罗里达大学的查尔斯·马丁教授的研究小组开发和论证。这项STTR工作将使Martin的团队与EIC实验室合作开发蛋白质组微阵列的新技术。为期12个月的I期研究旨在通过建立膜生产方法、证明潜在的光学成像现象、建立定量测定特征和变异系数以及证明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空白阵列和选定的MRA来商业化该技术。市场和产品类似于现有的蛋白质微阵列产品，新方法提供更快速和灵敏的检测。所提出的微阵列将应用于疾病检测/进展、蛋白质表达水平分析、小样品如活检的蛋白质分析、用于研究的蛋白质-蛋白质相互作用的鉴定、药物开发等，生物标志物发现这些应用已经指导了大量的蛋白质组微阵列市场;预计所提出的技术将在时间，成本和灵敏度方面对现有方法进行重大改进。

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.