Piezoelectric Pipetting for High Density Nucleic Acid Programmable Protein Arrays

描述(申请人提供)：在目前可用的高通量蛋白质组学技术中，蛋白质微阵列最有可能为分子诊断带来革命性的变化，用于早期检测、诊断、治疗、预后和监测临床反应。然而，由于制造的困难，蛋白质微阵列尚未充分发挥其作为研究或临床分子诊断工具的潜力。目前，蛋白质微阵列是通过表达和纯化数千种蛋白质来制造的，然后储存这些蛋白质，直到使用针式探测器打印出来，这一工艺流程存在许多固有的后勤问题。此外，许多蛋白质是不稳定的，所以这些步骤都必须保持在低温下。与PIN探测器相关的问题包括：相对较慢的打印速度、较差的斑点形态、PIN生物污垢问题、可变斑点大小、有限的微阵列密度等。因此，迫切需要更好、更便宜的蛋白质微阵列制造方法。在这笔赠款中，我们将结合两项成功的技术，开发一种大规模生产更快、更好、更便宜的蛋白质微阵列的创新方法。一种是基于我们先进的高速压电式移液管印制的DNA模板阵列，另一种是直接在微阵列表面原位表达蛋白质。工程艺术专业提供基于其专有的压电式移液技术的微阵列生产解决方案。LaBaer博士是核酸可编程蛋白质阵列(NAPPA)的共同发明者：这是第一种直接以微阵列形式原位表达蛋白质的方法。工程艺术公司将在亚利桑那州立大学生物设计研究所LaBaer博士的个性化诊断中心(CPD)安装其生产规模的压电微阵列机器(POC2)之一。我们将开发制造生产规模、商业级、高密度、可定制的蛋白质微阵列所需的工具、协议和过程控制，使其易于为广泛的蛋白质组学研究和临床诊断社区所用。这笔赠款直接呼吁开发一种广泛适用的研究工具，以应对蛋白质组学中的核心技术挑战。通过使高质量的蛋白质微阵列更易于评估，这笔赠款将有助于释放它们在研究和临床应用方面的真正潜力。这笔赠款将世界一流的压电移液管和工程艺术研发的电子产品结合在一起，后者在开发商业自动化生产规模的压电微阵列制造高密度全基因组基因表达微阵列方面拥有十多年的经验；来自新加坡一家老牌半导体生产设备制造商的世界级生产规模的自动化工艺制造设备、LaBaer博士获得专利的NAPPA技术，以及他在CPD中开发、表征和验证下一代商业蛋白质微阵列的能力。

DESCRIPTION (provided by applicant): Among the currently available techniques for high throughput proteomics, protein microarrays have the greatest prospects to revolutionize molecular diagnostics for early detection, diagnosis, treatment, prognosis and monitoring clinical response. However, protein microarrays have yet to reach their full potential as a research or clinical molecular diagnostics tool due to difficulties associated with their manufacture. Currently protein microarrays are manufactured by expressing & purifying thousands of proteins, which are then stored until they are printed using pin-spotters, a process flow with many inherent logistical problems. Furthermore, many proteins are unstable so these steps must all be maintained at cold temperature. Problems associated with pin spotters include: relatively slow printing speeds, poor spot morphology, pin biofouling issues, variable spot sizes, limited microarray densities and others. Thus, there are compelling needs for better and less expensive manufacturing methods for protein microarrays. In this grant we will combine two successful technologies to develop an innovative method for mass production of faster, better and cheaper protein microarrays. One technology is based on our advanced high speed piezoelectric pipettes to print arrays of cDNA templates and the other is to express proteins in situ directly on the microarray surface. Engineering Arts specializes in providing microarray production solutions based on its proprietary piezoelectric pipetting technology. Dr. LaBaer is the co-inventor of nucleic acid programmable protein arrays (NAPPA): the very first method to express proteins in situ directly in a microarray format. Engineering Arts will install one of its production-scale piezoelectric microarray machines (POC2) in Dr. LaBaer's Center for Personalized Diagnostics (CPD), Biodesign Institute, Arizona State University. We will develop tools, protocols and process controls required to manufacture production-scale, commercial-grade, high-density, customizable protein microarrays making them readily accessible to the broad proteomics research and clinical diagnostics communities. This grant directly addresses the call to develop a broadly applicable research tool that addresses a core technical challenge in proteomics. By making high quality protein microarrays more readily assessable, this grant will help unlock their true potential for research and clinical applications. This grant brings together world-class piezoelectric pipettes and electronics developed at Engineering Arts, over ten years experience in developing commercial automated production-scale piezoelectric microarraying manufacturing capabilities for high-density whole-genome gene expression microarrays; world class production-scale automation process manufacturing equipment from an established Singapore based semiconductor production equipment manufacturer, Dr. LaBaer's unique and patented NAPPA technology together in his CPD to develop, characterize and validate the next generation of commercial protein microarrays.