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

描述（由申请人提供）：拟议的研究是利用无喷嘴定向液滴喷射阵列和MEMS（微机电系统）技术，在固体底物上按需合成具有各种肽序列的二维蛋白质阵列。提出的研究是奠定了基础技术的便携式和负担得起的系统合成蛋白质探针阵列在固体基质上。蛋白质探针阵列合成系统是为了使蛋白质芯片技术适用于单个实验室，使实验室能够将数千个肽固定在固体底物上进行生物测定和筛选，并且在芯片设计上具有更大的灵活性，并且在制造新芯片时具有更快的周转时间。设想的便携式蛋白质合成系统基于无喷嘴定向喷射阵列和MEMS技术，可以使用少量蛋白质生成氨基酸在多孔或非多孔平面底物上快速简便地合成任何蛋白质序列的二维阵列。设想的“PECM”（蛋白质喷射芯片机）设计用于任何生物研究实验室；操作起来不会比标准的蛋白质合成器更困难。所提出的技术完全不同于在工厂生产预先制作的蛋白质探针的技术，也完全不同于预先制作蛋白质序列的微阵列技术。PECM使用新颖的微流体管理技术和自聚焦声学换能器阵列（SFAT）将20个蛋白质原氨基酸以任何所需的序列喷射到任何位置，以便在固体芯片上形成二维蛋白质探针阵列。PECM将是一个灵活、紧凑和低成本的系统，由二维SFAT喷射器阵列以及微流体组件、洗涤站、印刷电路板（包含控制和脉冲正弦波产生电路）、伺服电机和机械夹具组成，所有这些都集成在一个平台上。为了证明原理证明，一个由10 × 10组20个定向SFAT喷射器组成的阵列将与基于mems的微流体组件集成，用于在纤维素膜上产生100个任意指定序列的氨基酸蛋白质探针，这些探针具有适当的连接剂/间隔剂化学功能。膜上的单个位置将由20个定向SFAT喷射器喷墨，这些喷射器可以从垂直方向喷出任何方向的液滴，这样20个喷射器就可以在没有任何机械运动的情况下喷射器喷墨。因此，设想的PECM将有很少的运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底物上用二维阵列合成具有各种肽序列的蛋白质，使用定向喷射阵列根据需要喷射出纳米升的蛋白质原氨基酸液滴。这项研究将为便携式、灵活且价格合理的蛋白质合成系统奠定坚实的技术基础，该系统将在固体底物上生产任何蛋白质探针阵列（从少量的蛋白质生成氨基酸）。有了设想的蛋白质合成系统，蛋白质组学科学家将能够在进行生物分析时，以他们想要的任何方式在芯片上生成蛋白质探针（在几个小时内到达他们的位置）。

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.