Submerged Printing of Lipid and Membrane Protein Arrays

描述（由申请方提供）：该I期SBIR项目的目标是开发一种装置，用于将脂质和膜蛋白自动阵列打印到浸没的微量滴定板表面上，以保持其活性和功能。我们专有的3D微流体 印刷技术独特地能够以自动方式将生物分子阵列密封和印刷到浸没的表面上。通过在浸没的表面上打印，我们将产生脂质阵列，这些脂质阵列保持了细胞膜中发现的许多特征，并能够以多路复用的方式研究蛋白质-配体和蛋白质-膜相互作用。这样的工具将在包括癌症、糖尿病、炎症、感染和心血管疾病在内的众多领域中实现新型基于脂质的研究和诊断测定。Wasatch Microfluidics之前开发了一种基于流动的微流体打印技术，即连续流动微点仪（CFM），该技术使用3D通道网络，通过在离散点位置上来回流动将生物分子打印到平坦表面上。我们建议采取下一步，通过开发一个CFM打印头，可以打印在96孔微量滴定板的底部，适应CFM打印机硬件接受微量滴定板，并开发新的硬件，软件和技术，用于自动打印淹没表面。已经确定了以下具体目标，以证明使用Wasatch的CFM流式打印技术在96孔微量滴定板内浸没打印脂质和GPCR的可行性。1.设计一个CFM打印头，可以在标准96孔微量滴定板的孔内打印。2.调整CFM硬件和软件，以实现微量滴定威尔斯孔底部的自动浸没式打印。3.与Echelon Biosciences和大卫迈斯卡合作进行多重脂质阵列打印和分析。 公共卫生相关性：流体脂质双层和膜蛋白的组合构成生物细胞膜的主要结构组分。所有主要的人类疾病，包括代谢、自身免疫、血管、神经和癌症，都具有涉及功能失调的细胞和/或内膜的基本病理机制。通过使用目前可用的基于微量滴定板的技术对脂质和膜蛋白进行多重分析，有巨大的机会在其天然状态下研究这些系统，从而提高我们对受体结构和功能的认识。

DESCRIPTION (provided by applicant): The goal of this Phase I SBIR project is to develop a device for automated array printing of lipids and membrane proteins onto submerged microtiter plate surfaces in a way that maintains their activity and function. Our proprietary 3D microfluidic printing technology is uniquely capable of sealing and printing arrays of biomolecules onto submerged surfaces in an automated fashion. By printing onto submerged surfaces, we will produce lipid arrays that maintain many of the characteristics found in cellular membranes and enable the investigation of protein-ligand and protein-membrane interactions in a multiplexed fashion. Such a tool would enable novel lipid-based research and diagnostic assays in a multitude of areas including cancer, diabetes, inflammation, infections, and cardiovascular disease. Wasatch Microfluidics has previously developed a flow-based microfluidic printing technology, the Continuous Flow Microspotter (CFM), which uses 3D channel networks to print biomolecules onto flat surfaces by flowing them back and forth over discrete spot locations. We propose to take the next step by developing a CFM printhead that can print within the bottom of a 96 well microtiter plate, adapting the CFM printer hardware to accept microtiter plates, and developing new hardware, software and techniques for automated printing on submerged surfaces. The following specific aims have been identified to prove the feasibility of using Wasatch's CFM flow printing technology for submerged printing of lipids and GPCRs within 96 well microtiter plates. 1. Design a CFM printhead that can print within the well of a standard 96 well microtiter plate. 2. Adapt the CFM hardware and software to enable automated submerged printing on the bottom of microtiter wells. 3. Multiplex lipid array printing and analysis in partnership with Echelon Biosciences and David Myszka. PUBLIC HEALTH RELEVANCE: The combination of fluid lipid bilayers and membrane proteins constitute the major structural components of biological cell membranes. All major human diseases including metabolic, autoimmune, vascular, neurological, and cancer have essential pathologic mechanisms involving dysfunctional cell and/or internal membranes. By enabling multiplexed analysis of lipids and membrane proteins using currently available microtiter plate based technologies, there is a tremendous opportunity to study these systems in their native states and thereby improve our knowledge of receptor structures and functions.