Piezoelectric Pipetting for High Density Nucleic Acid Programmable Protein Arrays

用于高密度核酸可编程蛋白质阵列的压电移液

• 批准号: 8001744
• 负责人: PETER J. WIKTOR
• 金额: $ 21.33万
• 依托单位: ENGINEERING ARTS, LLC
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8001744
• 关键词: Address; Amines; Antibodies; Arizona; Arts; Asses; Autoantibodies; Automation; Binding; Biochemistry; Biological Assay; Biological Markers; Biology; Bovine Serum Albumin; Chemistry; Clinical; Communities; Community Healthcare; Complementary DNA; DNA; DNA Binding; Deposition; Detection; Development; Diagnosis; Diagnostic; Diffusion; Disease; Dose; Early Diagnosis; Electronics; Elements; Engineering; Enzyme-Linked Immunosorbent Assay; Equipment; Fluorocarbons; Functional disorder; Gene Expression; Genes; Genetic Transcription; Genome; Glass; Goals; Grant; Head; Health Services Research; Hour; Human; Human Resources; Image; Immunoassay; In Situ; In Vitro; Individual; Industry; Inherited; Institutes; Legal patent; Libraries; Location; Logistics; MDM2 gene; Manufacturer Name; Measurement; Mechanics; Methods; Metric; Microfabrication; Molecular; Monitor; Monoclonal Antibodies; Morphology; Nucleic Acids; Performance; Phase; Plasmids; Printing; Process; Production; Protein Array; Protein Microchips; Protein p53; Proteins; Proteomics; Protocols documentation; Publishing; Quality Control; Recovery; Relative (related person); Reproducibility; Research; Resolution; Sampling; Screening Result; Screening procedure; Semiconductors; Serum; Signal Transduction; Signaling Protein; Silicon; Silicon Dioxide; Singapore; Slide; Small Business Technology Transfer Research; Solutions; Specific qualifier value; Speed; Spottings; Surface; Surface Plasmon Resonance; System; TP53 gene; Techniques; Technology; Temperature; Testing; Therapeutic; Time; Titrations; Translations; Universities; Variant; Western Blotting; Work; base; cDNA Arrays; chemical reaction; clinical application; cold temperature; cost; cross reactivity; density; experience; improved; innovation; interdisciplinary approach; manufacturing process; miniaturize; monolayer; nanolitre; next generation; novel; novel strategies; operation; outcome forecast; polyclonal antibody; prevent; protein function; protein protein interaction; prototype; public health relevance; quality assurance; response; success; technological innovation; tool; vector

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. PUBLIC HEALTH RELEVANCE: Nearly all diagnostics and therapeutics act through proteins, which are the working machines of biology. The study of proteins, both their activities and their dysfunction in disease, has been historically managed one- protein-at-a-time; however, this will be dramatically accelerated through the use of protein microarrays, which microscopically display thousands of functional proteins. This grant will develop technology to mass produce better and less expensive protein microarrays, making them more readily accessible to the broad research and health care communities.

描述（由申请人提供）：在目前可用的高通量蛋白质组学技术中，蛋白质微阵列在分子诊断的早期检测、诊断、治疗、预后和临床反应监测方面具有最大的前景。然而，由于制造方面的困难，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，然后将这些蛋白质储存起来，直到使用pin-spotters进行打印，这是一个具有许多固有后勤问题的工艺流程。此外，许多蛋白质是不稳定的，所以这些步骤都必须在低温下进行。与引脚点相关的问题包括：相对较慢的打印速度、较差的点形态、引脚生物污垢问题、可变的点尺寸、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。在这笔拨款中，我们将结合两项成功的技术，开发一种创新的方法，用于更快、更好、更便宜的蛋白质微阵列的大规模生产。一种技术是基于我们先进的高速压电移液器来打印cDNA模板阵列，另一种技术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阵列（NAPPA）的共同发明者：这是第一种直接在微阵列格式中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）。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，使其易于广泛的蛋白质组学研究和临床诊断社区使用。这项资助直接用于开发一种广泛适用的研究工具，以解决蛋白质组学的核心技术挑战。通过使高质量的蛋白质微阵列更容易评估，这笔拨款将有助于释放它们在研究和临床应用方面的真正潜力。这项资助汇集了世界级的压电移液器和工程艺术开发的电子产品，在开发高密度全基因组基因表达微阵列的商业自动化生产规模的压电微阵列制造能力方面超过十年的经验；来自新加坡半导体生产设备制造商的世界级生产规模自动化工艺制造设备，LaBaer博士独特的专利NAPPA技术在他的CPD中一起开发，表征和验证下一代商业蛋白质微阵列。