An Integrated Microarray Printing and Detection System

 描述（由申请人提供）：微阵列技术极大地推进了蛋白质相互作用的研究，从而发现和验证了新的生物标志物和治疗药物。通常，蛋白质微阵列在一个装置中预先点样靶分子，然后使用另一个装置用探针分子进行测试。在每个测试周期中，整个微阵列暴露于探针分子。尽管这种方法已经被证明对于某些应用是有效的，但是它固有地受到阻碍其获得更广泛实用性的几个关键限制：1）打印和分析步骤都消耗大量样品体积，这在只有少量蛋白质可用或负担得起的情况下尤其成问题; 2）样品溶液暴露于整个微阵列限制了仅一个探针与N个靶的动力学相互作用分析（仅1XN相互作用），显著限制了微阵列的应用类型和分析能力;和3）完整的微阵列必须盲式预打印，而没有关于点均匀性，靶活性，或探针选择性，这可能导致不确定的数据、不必要的测试和获得有效结果的延迟。我们提出了一个集成的微阵列打印和检测系统（IMPDS），以解决限制分析能力和更广泛的吸引力的微阵列技术的关键限制。IMPDS将有能力纳入微阵列形成和测试的反馈，以便更快地产生更多相关结果，用超低纳升体积的样品进行高分辨率基于液滴的测试，对高密度微阵列进行更通用的MxN（多对多）蛋白质动力学相互作用分析，并测量基于细胞的微阵列中的分子相互作用和结合动力学。IMPDS依赖于将两项核心技术精心集成到一台仪器中：1）新型超低容量压电液体分配系统和2）专有的高分辨率无失真表面等离子体共振成像（DF-SPRi）系统。该项目的成功将导致一种新的可商业化的微阵列技术，该技术能够：1）将微阵列点样和检测简化到单个仪器中，以获得更简单、更快速、更准确的结果，2）高密度微阵列的基于超低体积纳米液滴的分析，3）真实的时间内的灵活和多重M × N无标记蛋白质相互作用动力学分析，4）具有单细胞分辨率的基于细胞的微阵列分析。该项目将汇集生物传感仪器公司的优势。(BI)来自亚利桑那州州立大学（ASU）生物设计研究所的生物电子和生物传感器中心，是压电液体点胶技术的发明者。我们将共同开发IMPDS成为高通量蛋白质相互作用研究中商业用途的强大工具，从而发现和验证新的分子诊断生物标志物和新的治疗药物。

 DESCRIPTION (provided by applicant): Microarray technology has dramatically advanced the study of protein interactions leading to discovery and validation of new biomarkers and therapeutic drugs. Typically, protein microarrays are pre-spotted with target molecules in one device and then tested with probe molecules using another device. During each testing cycle, the entire microarray becomes exposed to the probe molecules. Though this approach has been shown to be effective for some applications, it inherently suffers from several key limitations that hinder it from gaining broader utility: 1) both the printing and analysis steps consume large sample volumes, which is particularly problematic where only small amounts of proteins are available or affordable; 2) the exposure of sample solution to the entire microarray restricts the kinetic interaction analysis of only one probe to N targets (1XN interactions only), significantly limiting the types of applications and analytical power of microarrays; and 3) complete microarrays must be pre-printed blindly with no feedback on spot uniformity, target activity, or probe selectivity which may lead to inconclusive data, unnecessary tests, and delays in obtaining effective results. We propose an Integrated Microarray Printing and Detection System (IMPDS) to address the key limitations restricting the analytical power and broader appeal of microarray technology. IMPDS will have the ability to incorporate feedback of microarray formation and testing in order to generate more relevant results sooner, perform high resolution droplet-based testing with ultra-low nanoliter volume samples, conduct a more versatile M x N (many-to-many) protein kinetic interaction analysis of high density microarrays, and measure molecular interactions and binding kinetics in cell-based microarrays. IMPDS relies upon the careful integration of two core technologies into a single instrument: 1) a novel ultra-low volume piezoelectric liquid dispensing system and 2) a proprietary, high-resolution, distortion-free surface plasmon resonance imaging (DF-SPRi) system. The success of this project will lead to a new commercializable microarray technology capable of: 1) streamlining microarray spotting and detection into a single instrument for simpler, faster, more accurate results, 2) ultra-low volume nanodroplet-based analysis of high density microarrays, 3) flexible and multiplexed M x N label-free protein interaction kinetic analyses in real time, 4) cell-based microarray analyses with single cell resolution. This project will bring together strengths from Biosensing Instrument Inc. (BI) an innovator and global supplier of high-performance SPR instruments, and from the Center for Bioelectronics and Biosensors, the Biodesign Institute at Arizona State University (ASU), inventors of the piezoelectric liquid dispensing technology. Together we will develop IMPDS into a powerful tool for commercial use in high-throughput protein interaction studies leading to the discovery and validation of new molecular diagnostic biomarkers and new therapeutic drugs.