DEVELOPMENT OF AN UNLABELED MACROMOLECULE DETECTOR

描述（由申请人提供）：本研究的总体目标是开发一种用于功能基因组学和系统生物学的仪器，能够识别和定量复杂混合物中的未标记大分子（蛋白质和核酸）。 该系统将以高度平行的检测格式在单分子检测模式中识别大分子，从而避免与系综测量相关的信息丢失，同时实现统计学上显著的样本量以避免假结果。 该系统将与三种不同的测量设置一起使用：溶液中的大分子、细胞表面的大分子和固定化生物化学平台。 该仪器基于检测大分子之间的相互作用和新发明的光子效应SPEI。 SPEI是一种光子学效应，能够通过亚波长孔径阵列实现高的光透射系数。 目前制造的这些孔的直径为50 nm，预计直径小至10 nm是可能的。 该仪器开发所基于的假设是，大分子将通过改变发射光的强度或改变阵列的共振来改变来自孔阵列的光的发射。 这些效应将产生独特的特征，其可用于在单分子水平上以统计学显著的样本量检测和定量大分子。 在R21阶段的研究将集中在展示这些大分子与SPEI相互作用的影响，并产生/识别特征签名。 R33阶段的研究将集中在开发一种仪器系统，该系统可以控制大分子通过孔的流动，并操纵生物化学和物理环境。 然后，该系统将被应用于溶液中的大分子，细胞表面现象和固定化生物化学（例如蛋白质微阵列）的研究。

DESCRIPTION (provided by applicant): The overall objective of this research is to develop an instrument for use in functional genomics and system biology that is capable of identifying and quantifying unlabeled macromolecules (proteins and nucleic acids) in complex mixtures. This system will identify the macromolecules in single molecule detection mode with a highly parallel detection format, thereby avoiding the loss of information associated with ensemble measurements while achieving statistically significant sample sizes to avoid spurious results. This system will be used with three different measurement settings: macromolecules in solution, macromolecules at the cell surface, and immobilized biochemistry platforms. This instrument is based on detecting interactions between the macromolecules and a newly invented photonics effect, SPEI. SPEI is a photonics effect that is capable to achieving high light transmission coefficients through an array of subwavelength apertures. These apertures are currently being fabricated with diameters of 50 nm and it is expected that diameters as small as 10 nm are possible. The hypothesis upon which this instrument development is based is that macromolecules will alter the emission of light from the array of apertures by changing the intensity of the emitted light or shifting the resonance of the array. These effects will create a unique signature that can be used to detect and quantify macromolecules at the single molecule level with statistically significant sample sizes. The research in the R21 phase will be focused on demonstrating the effects of these macromolecular interactions with SPEI and generating/identifying characteristic signatures. The research in the R33 phase will be focused on developing an instrument system that can control the flow of the macromolecules through the apertures and manipulate the biochemical and physical environments. This system will then be applied to the study of macromolecules in solution, cell surface phenomena, and immobilized biochemistry (e.g. protein microarrays).