An Integrated Microarray Printing and Detection System

 描述(申请人提供)：微阵列技术极大地促进了蛋白质相互作用的研究，导致了新的生物标记物和治疗药物的发现和验证。通常，蛋白质微阵列在一个设备中预先发现目标分子，然后使用另一个设备与探针分子进行测试。在每个测试周期中，整个微阵列都会暴露在探针分子中。尽管这种方法已被证明对某些应用是有效的，但它固有的几个关键限制阻碍了它获得更广泛的实用：1)打印和分析步骤都消耗大量样品，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)样品溶液暴露在整个微阵列中限制了只有一个探针与N个靶的动力学相互作用分析(仅1×N个相互作用)，大大限制了微阵列的应用类型和分析能力；3)完整的微阵列必须盲目预印，没有关于斑点一致性、靶标活性或探针选择性的反馈，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。我们提出了一种集成的微阵列打印和检测系统(IMPDS)，以解决限制微阵列技术分析能力和更广泛吸引力的关键限制。IMPDS将能够结合微阵列形成和测试的反馈，以便更快地产生更相关的结果，使用超低纳升体积的样品执行基于液滴的高分辨率测试，对高密度微阵列进行更多功能的M x N(多对多)蛋白质动力学相互作用分析，以及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分配系统和2)专有的高分辨率、无失真表面等离子共振成像(DF-SPRI)系统。该项目的成功将带来一种新的可商业化的微阵列技术，能够：1)将微阵列检测和检测简化到单一仪器中，以实现更简单、更快、更准确的结果；2)基于超低容量纳米液滴的高密度微阵列分析；3)灵活和多路复用的M x N无标记蛋白质相互作用实时动力学分析；4)单细胞分辨率的基于细胞的微阵列分析。这个项目将汇集高性能SPR仪器的创新者和全球供应商Biosensing Inc.(BI)以及压电液分配技术的发明者、生物电子和生物传感器中心、亚利桑那州立大学(ASU)生物设计研究所的力量。我们将共同将IMPDS发展成为高通量蛋白质相互作用研究中用于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记物和新的治疗药物。

 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.