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: 7708805
• 负责人: EUN SOK KIM
• 金额: $ 23.93万
• 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 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7708805
• 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）：拟议的研究是使用在固体基材上综合二维蛋白质，使用固体基板上的各种肽序列，使用小型，定向的液滴 - 注射器阵列和MEMS和MEMS（Microelectromechorical Systems）技术。拟议的研究是为固体基材上蛋白质探针阵列合成便携式和负担得起的系统的基础技术。蛋白质探针阵列的合成系统是为了使蛋白质芯片技术可用于单个实验室，以便实验室能够将数千种肽固定在固体基板上进行生物测定和筛查，并在芯片设计方面具有更大的灵活性，并在制造新的碎屑方面更快地旋转。设想的便携式蛋白质合成系统基于一个无Zimental的弹出器阵列和MEMS技术，以实现使用少量蛋白质激氨基酸的多孔或非孔平面底物上任何蛋白质序列的二维阵列的快速合成。设想的“ PECM”（蛋白质喷射芯片机）旨在用于任何生物学研究实验室；与标准蛋白质合成器相比，操作不会更困难。所提出的技术与在工厂预先制作的蛋白质探针的技术完全不同，并且与发现预制蛋白质序列的微阵列技术也完全不同。 PECM使用新型的微流体管理技术和一系列自我关注的声音传感器（SFAT）以任何所需的序列将20个蛋白质氨基酸驱射到任何位置，以形成固体芯片上的2-D蛋白质探针。 PECM将是一个灵活，紧凑且低成本的系统，由2D SFAT弹出器以及微流体组件，一个洗涤站，印刷电路板（包含控制和脉冲 - 鞋底波电通路），伺服机和机械固定装置以及所有单个平台上的机械装置组成。为了证明原理证明，将与基于MEMS的微流体成分集成10 x 10组的阵列，该阵列将集成，用于在纤维素膜上与适当的接头/SPACER化学功能化的纤维素膜上任意指定序列的氨基酸的100个蛋白质探针。膜上的一个位置将由20个方向弹出器墨水墨水，该液滴可以从直垂直方向沿任何方向弹出液滴，以便可以通过20个弹出器将斑点墨水墨水，而无需弹出器的任何机械运动。因此，设想的PECM将几乎没有运动部件和非严格的对齐要求。公共卫生相关性：拟议的研究将使用定向喷射器阵列在固体底物上具有各种肽序列的蛋白质的2-D阵列合成，以按需驱射蛋白质氨基酸的纳米液滴。这项研究是为便携式，柔性且负担得起的蛋白质合成系统奠定牢固的技术基础，该基础将在固体底物上产生任何蛋白质探针阵列（来自少量的蛋白质氨基酸）。借助设想的蛋白质合成系统，蛋白质组学科学家将能够以任何方式（在数小时之内）以任何方式在芯片上生成蛋白质探针，因为他们进行了生物测定。