On-Chip Protein Synthesis Based on Directional Droplet-Ejector Array

基于定向液滴喷射器阵列的芯片上蛋白质合成

• 批准号: 7938577
• 负责人: EUN SOK KIM
• 金额: $ 19.85万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7938577
• 关键词: Acoustics; Affect; Amino Acid Sequence; Amino Acids; Antibodies; Binding; Biological Assay; Biomedical Engineering; Caliber; Cellulose; Chemistry; Coupling; DNA biosynthesis; Deposition; Effectiveness; Foundations; Generations; Goals; Head; Heating; Hour; Individual; Ink; Laboratories; Length; Location; Mechanics; Membrane; Microfluidics; Motion; Noise; Peptide Sequence Determination; Peptides; Physiologic pulse; Printing; Protein Array; Protein Biosynthesis; Protein Microchips; Proteins; Proteomics; Reproducibility; Research; Research Proposals; Scientist; Screening procedure; Side; Signal Transduction; Site; Solid; Specific qualifier value; Spottings; System; Techniques; Technology; Time; Transducers; base; biological research; cost; design; flexibility; high risk; nanolitre; novel; protein aminoacid sequence; public health relevance; scale up; two-dimensional

DESCRIPTION (provided by applicant): The proposed research is to synthesize a 2-D array of proteins with various peptide sequences on a solid substrate on demand, using a nozzleless, directional droplet-ejector array and MEMS (microelectromechanical systems) technology. The proposed research is to lay down the underlying technologies for a portable and affordable system for synthesis of protein probe arrays on a solid substrate. The synthesis system for protein probe array is to make protein chip technology available to individual laboratories so that the laboratories may be able to immobilize thousands of peptides on a solid substrate for bioassay and screening, and have far greater flexibility in chip design and faster turnaround in fabricating new chips. The envisioned portable protein synthesis system is based on a nozzleless, directional ejector array and MEMS technology to enable the rapid and facile synthesis of a two-dimensional array of any protein sequence on a porous or nonporous planar substrate using small quantities of proteinogenic amino acids. The envisioned "PECM" (Protein Ejector Chip Machine) is designed for use in any biological research laboratory; it will be no more difficult to operate than a standard protein synthesizer. The proposed technique is entirely different from the technology that produces protein probes pre-made at factory, and completely different also from microarray techniques that spot pre-made protein sequences. The PECM uses novel microfluidic management techniques and an array of self-focusing acoustic transducer (SFAT) for ejecting the 20 proteinogenic amino acids in any desired sequence to any location in order to form a 2-D array of protein probes on a solid chip. The PECM will be a flexible, compact and low cost system that consists of a 2-D array of SFAT ejectors along with microfluidic components, a wash station, a printed circuit board (that contains the control and pulsed-sinusoidal-wave-generation circuits), servomotors and mechanical fixtures, all integrated on a single platform. To demonstrate proof-of-principle, an array of 10 x 10 sets of 20 directional SFAT ejectors will be integrated with MEMS-based microfluidic components for producing 100 protein probes of amino acids of arbitrarily specified sequence on a cellulose membrane functionalized with appropriate linker/spacer chemistry. A single location on the membrane will be inked by 20 directional SFAT ejectors that can eject droplets in any direction off from the straight vertical direction so that a spot can be inked by 20 ejectors without any mechanical motion of the ejectors. Consequently, the envisioned PECM will have few moving parts and non- stringent alignment requirements. PUBLIC HEALTH RELEVANCE: The proposed research will demonstrate 2-D array synthesis of proteins with various peptide sequences on a solid substrate, using a directional ejector array to eject nanoliter droplets of proteinogenic amino acids on demand. The research is to lay down a firm technical foundation for a portable, flexible and affordable protein synthesis system that will produce any protein probe array on a solid substrate (from small amounts of proteinogenic amino acids). With the envisioned protein synthesis system, proteomics scientists will be able to generate protein probes on a chip in any way they desire (at their sites within hours), as they carry out bioassays.

描述（由申请人提供）：拟议的研究是根据需要在固体基质上合成具有各种肽序列的蛋白质的2-D阵列，使用无毛刺的定向液滴喷射器阵列和MEMS（微机电系统）技术。该研究旨在为一种便携式和负担得起的系统奠定基础技术，用于在固体基底上合成蛋白质探针阵列。蛋白质探针阵列的合成系统是使蛋白质芯片技术可用于各个实验室，以便实验室能够将数千个肽段固定在固体基底上进行生物测定和筛选，并且在芯片设计方面具有更大的灵活性，并且在制造新芯片方面具有更快的周转时间。设想的便携式蛋白质合成系统是基于一个无毛刺的，定向喷射器阵列和MEMS技术，使任何蛋白质序列的二维阵列的多孔或无孔平面基板上使用少量的蛋白质氨基酸的快速和简便的合成。设想中的“PECM”（蛋白质喷射芯片机）是为任何生物研究实验室设计的;它不会比标准蛋白质合成仪更难操作。所提出的技术与在工厂生产预制蛋白质探针的技术完全不同，也与发现预制蛋白质序列的微阵列技术完全不同。PECM使用新的微流体管理技术和自聚焦声换能器（SFAT）阵列，用于以任何期望的序列将20种蛋白质氨基酸喷射到任何位置，以便在固体芯片上形成蛋白质探针的2-D阵列。PECM将是一个灵活、紧凑和低成本的系统，包括一个二维阵列的SFAT喷射器沿着与微流体组件、清洗站、印刷电路板（包含控制和脉冲正弦波发生电路）、伺服电机和机械夹具，所有这些都集成在一个平台上。为了证明原理，将10 x 10组20个定向SFAT喷射器的阵列与基于MEMS的微流体组件集成，用于在用适当的连接体/间隔物化学功能化的纤维素膜上产生100个任意指定序列的氨基酸的蛋白质探针。膜上的单个位置将由20个定向SFAT喷射器上墨，所述喷射器可以在偏离直线垂直方向的任何方向上喷射液滴，使得点可以由20个喷射器上墨，而无需喷射器的任何机械运动。因此，设想的PECM将具有很少的移动部件和非严格的对准要求。公共卫生相关性：拟议的研究将展示在固体基底上具有各种肽序列的蛋白质的2-D阵列合成，使用定向喷射器阵列根据需要喷射蛋白质氨基酸的纳升液滴。这项研究将为一个便携、灵活和负担得起的蛋白质合成系统奠定坚实的技术基础，该系统将在固体基质上（从少量蛋白质氨基酸）生产任何蛋白质探针阵列。有了设想的蛋白质合成系统，蛋白质组学科学家将能够在芯片上以任何他们想要的方式（在几个小时内在他们的地点）生成蛋白质探针，因为他们进行生物测定。