Membrane Receptor Microarrays Based on Quantum Dot Barcoded Lipobeads

摘要建议标题：基于量子点条形码脂珠的膜受体微阵列，建议编号：CTS-0626139，首席研究员：Alexander Couzis，机构：纽约市立大学摘要：本项目将开发一种新的用于显示表面细胞膜受体的微阵列平台。该阵列旨在用于高通量筛选膜受体与潜在结合伙伴的结合。在阵列设计中，受体被隔离在一个支撑的双层中，该双层包裹着直径1m的条形码二氧化硅或聚苯乙烯微珠形成脂珠，在双层中插入膜受体使受体保持其生物活性。通过将脂珠插入微米大小的孔(略大于脂珠)，将脂珠排列在表面上。微孔在表面上以方形网格形成图案，其中孔以3m的中心到中心间隔隔开。使用嵌入到颗粒中的发光量子点制成的标签对封装的颗粒进行条形化，并共价连接到二氧化硅或聚苯乙烯基质。该阵列在1 mm x 1 mm的有效区域上有大约105个注册表，每个磁珠注册表能够显示不同的受体。检测结合事件，并使用荧光显微镜读取识别脂珠上受体的条形码。该阵列将通过筛选细胞膜隔离的糖脂与细菌毒素的相互作用来进行测试。该项目的成功将对蛋白质的微阵列，特别是膜受体的微阵列产生重大影响：该平台允许小型化，比任何现有的膜受体或蛋白质展示平台小一个数量级，并提供对膜受体的严格要求的膜受体的简单展示。这种阵列将提供更强的探测膜受体结合相互作用的能力，这对药物发现特别重要，因为药物的分子靶标有50%是膜结合的。它也可以被证明是一种重要的工具，用于定位和理解细胞蛋白质组在高达25%的给定生物膜蛋白密码子的序列基因中的结合作用。从更广泛的影响角度来看，该提案广泛涉及拟议工作的潜在经济和社会效益。PIs将参加CCNY的高中外展计划。该项目将包括一名研究生和本科生的培训，该项目将与PI合作，并通过CCNY现有的项目提供支持。PI已经证明了她可以很好地与本科生合作，这是拟议工作的一个有价值的方面。因此，该提案的教育方面是该提案的一个优点。

Abstract Proposal Title: Membrane Receptor Microarrays Based on Quantum Dot Barcoded Lipobeads, Proposal Number: CTS-0626139, Principal Investigator: Alexander Couzis, Institution: City University of New York Abstract:A new microarray platform for the display of surface cell membrane receptors is will be developed in this project. The array is intended to be used for the high throughput screening of the binding of membrane receptors with potential binding partners. In the array design, the receptors are sequestered in a supported bilayer, which encapsulates a 1 m in diameter barcoded silica or polystyrene bead to form a lipobead.Insertion of the membrane receptor in the bilayer allows the receptor to retain its biological activity. The lipobeads will be arrayed on a surface by inserting them intomicron sized wells (slightly larger than the lipobead). The micro-wells are patterned in asquare grid on the surface in which wells are separated by a center-to-center spacing of3m. The encapsulated particles are barcoded using a label made of luminescent quantumdots embedded into the particles, and covalently linked to the silica or polystyrenematrix. The array has approximately 105 registries on a 1mm x 1mm active area, with thelipobead registries capable of each displaying a different receptor. Binding events aredetected and the barcode identifying the receptor on the lipobead read using fluorescencemicroscopy. The array will be tested by screening the interaction of a cell membrane sequestered glycolipid with a bacterial toxin. Preliminary results are presented on the synthesis of the quantum dot encoded beads and the fabrication of the micro-well patterned surface.The success of this program will have a significant impact in the microarraying of proteins in general and membrane receptors in particular: The platform allows for a miniaturization that is orders of magnitude smaller than any current platforms for the display of membrane receptors or proteins in general, and provides for the facile display of difficult to exhibit membrane receptors with their strict requirement of a lipid environment. The enhanced ability to probe the binding interactions of membrane receptors which this array will afford is particularly important to drug discovery since fifty percent of the molecular targets of drugs are membrane bound. It could also prove be an important tool for the mapping and understanding of the binding interactions of the cell proteome in as much as twenty five percent of the sequenced genes of a given organism code for membrane proteins. From a broader impacts perspective, the proposal broadly addresses the potential economic and societal benefits of the proposed work. The PIs will participate in CCNY's high school outreach program. The project would include the training of a graduate student and undergraduates will work with the PI and be supported through pre-existing programs at CCNY. The PI has demonstrated that she can work well with undergraduate students, which is a valuable aspect of the proposed work. The education aspects of the proposal are thus a strength of the proposal.