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: 8716785
• 负责人: PETER J. WIKTOR
• 金额: $ 59.96万
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8716785
• 关键词: Address; Amines; Antibodies; Arizona; Arts; Asses; Autoantibodies; Automation; Beryllium; 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; Engineering; Enzyme-Linked Immunosorbent Assay; Equipment; Fluorocarbons; Functional disorder; Gene Chips; 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; 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; 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; quality assurance; response; screening; 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.

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，蛋白质微阵列最有可能彻底改变分子诊断，以实现早期检测、诊断、治疗、预后和监测临床反应。然而，由于制造困难，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，直到使用针点样机进行打印，这是一个存在许多固有后勤问题的工艺流程。此外，许多蛋白质不稳定，因此这些步骤都必须保持在低温下。与针点样器相关的问题包括：相对较慢的打印速度、较差的点形态、针生物污染问题、可变的点尺寸、有限的微阵列密度等。因此，迫切需要更好且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来开发一种创新方法，用于大规模生产更快、更好和更便宜的蛋白质微阵列。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。 Engineering Arts 专注于提供基于其专有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质阵列 (NAPPA) 的共同发明人：这是第一种以微阵列形式直接原位表达蛋白质的方法。 Engineering Arts 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) 安装一台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，使广泛的蛋白质组学研究和临床诊断社区可以轻松获得它们。这笔赠款直接响应了开发一种广泛适用的研究工具的号召，以解决蛋白质组学的核心技术挑战。通过使高质量蛋白质微阵列更容易评估，这笔赠款将有助于释放其研究和临床应用的真正潜力。这笔资助汇集了 Engineering Arts 开发的世界一流的压电移液器和电子产品，以及在开发用于高密度全基因组基因表达微阵列的商业自动化生产规模压电微阵列制造能力方面十多年的经验； LaBaer 博士将来自新加坡知名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与独特的 NAPPA 专利技术结合到他的 CPD 中，以开发、表征和验证下一代商业蛋白质微阵列。

项目成果

Microreactor array device.

• DOI: 10.1038/srep08736
• 发表时间: 2015-03-04
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Wiktor P;Brunner A;Kahn P;Qiu J;Magee M;Bian X;Karthikeyan K;LaBaer J
• 通讯作者: LaBaer J

Antiviral antibody profiling by high-density protein arrays.

• DOI: 10.1002/pmic.201400612
• 发表时间: 2015-06
• 期刊: Proteomics
• 影响因子: 3.4
• 作者: Bian X;Wiktor P;Kahn P;Brunner A;Khela A;Karthikeyan K;Barker K;Yu X;Magee M;Wasserfall CH;Gibson D;Rooney ME;Qiu J;LaBaer J
• 通讯作者: LaBaer J