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

描述(申请人提供)：拟议的研究是利用无喷嘴、定向液滴喷射器阵列和MEMS(微型机电系统)技术，在固体基质上按需合成具有各种肽序列的二维蛋白质阵列。这项拟议的研究是为在固体基质上合成蛋白质探针阵列的便携式和负担得起的系统奠定基础技术。蛋白质探针阵列的合成系统是将蛋白质芯片技术提供给各个实验室，使实验室能够将数千个多肽固定在固体底物上进行生物检测和筛选，并在芯片设计方面具有更大的灵活性，在制造新芯片时更快地周转。设想的便携式蛋白质合成系统基于无喷嘴定向喷射器阵列和MEMS技术，能够使用少量的蛋白质生成氨基酸在多孔或非多孔平面底物上快速而简便地合成任意蛋白质序列的二维阵列。设想中的“PECM”(蛋白质喷射器芯片机)设计用于任何生物研究实验室；它的操作不会比标准蛋白质合成器更难操作。这项拟议的技术与生产工厂预制的蛋白质探针的技术完全不同，也与检测预制蛋白质序列的微阵列技术完全不同。PECM使用新颖的微流控管理技术和一组自聚焦声学换能器(SFAT)，将所需序列中的20种蛋白质原氨基酸喷射到任何位置，以便在固体芯片上形成2-D蛋白质探针阵列。PECM将是一个灵活、紧凑和低成本的系统，由SFAT喷射器的2-D阵列以及微流控部件、洗涤工位、印刷电路板(其中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。为了演示原理证明，一个由10×10组20个定向SFAT喷射器组成的阵列将与基于MEMS的微流控部件集成在一起，在具有适当连接物/间隔物化学功能的纤维素膜上生产100个任意指定序列的氨基酸蛋白质探针。膜上的单个位置将由20个方向的SFAT喷射器进行喷墨，这些喷射器可以从垂直直线方向向任何方向喷射液滴，因此可以由20个喷射器对一个斑点进行喷墨，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不严格的对准要求。与公共健康相关：拟议的研究将展示在固体底物上合成具有不同肽序列的蛋白质的二维阵列，使用定向喷射器阵列根据需要喷射纳升蛋白质产生的氨基酸液滴。这项研究旨在为便携式、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(从少量的蛋白质产生氨基酸)产生任何蛋白质探针阵列。有了设想中的蛋白质合成系统，蛋白质组学科学家将能够在芯片上以他们想要的任何方式产生蛋白质探针(在几小时内在他们的位置上)，同时进行生物检测。

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.