BioMarker Analysis from Single Cells using Nanobiosensor

描述（由申请人提供）：本I期可行性研究的目的是设计、构建和评价一种超小型传感器装置，该装置能够检测来自少数细胞和最终单细胞的ng蛋白质生物标志物。该装置被称为纳米生物传感器，由一系列超小型化的蛋白质捕获（抗体）域印刷在一个微制造的硅结构。纳米生物传感器结合了纳米技术、细胞生物学和分子生物学的概念、方法和仪器，为单细胞蛋白质组学分析提供了一条途径。纳米生物传感器将用于监测模型细胞系统中分泌和细胞内蛋白质的表达。纳米生物传感器的设计允许将测试直接带到样品来源（在这种情况下，细胞），消除了与将极少量样品提取物运输到远程测试地点相关的困难。本发明通过申请机构开发的“纳米阵列”技术来实现，该技术用于构建具有超微（1-20 mm）至亚微米（250 nm）空间地址（对应于常规微阵列测试所需表面积的1/1000 - 1/10，000）并且仅需要极小样品体积（亚微升至皮升）的生物芯片。纳米生物传感器将允许分析来自样品的蛋白质谱，以前只能通过PCR扩增核酸进行分析。所提出的纳米生物传感器打开了快速和信息分析微小蛋白质样品的大门，例如在法医情况下，新生儿和组织亚群（例如，激光细胞显微切割活检样本）。这种微型化水平转化为微创但数据丰富的分子诊断，用于癌症和其他疾病的早期检测，显著减少患者的不适。

DESCRIPTION (provided by applicant): The purpose of this Phase I feasibility study to design, construct and evaluate an ultraminiaturized sensor device capable of detect ng protein biomarkers from a few cells and, ultimately, single cells. The device, termed the nanobiosensor, consists of an array of ultraminiaturized protein captur e (antibody) domains printed on a micro fabricated silicon structure. The nanobiosensor combines concepts, methods and instrumentation from nanotechnology and cel I and molecular biology to create a pathway to single-cell proteomic analysis. The nanobiosensor will be used for monitoring both secreted and intracellular protein expression in model cell ular systems. The design of the nanobiosensor allow s the test to be brought directly to the sample origin (in this case, a cell), eliminating the difficulties associated with transporting extremely small quantities of sample extracts to a remote test site. The pr opposed research is enabled by "NanoArrayer" technology developed by the applicant institution for the construction of biochips w itch ultramicro (1-20 mm) to submicron (250 nm) spatial addresses (corresponding to 1/1000th - 1/10,000th of the surface area required for conventional microar rays tests) and requiring only extremely small sample volumes (sub-microliter to picoliter). The nanobiosensor will permit analysis of protein profiles from samples that previously could only be analyzed via PCR amplification of nucleic acids. The proposed nanobiosensor opens the door to fast and infor mative analysis of minute protein samples such as those found in forensics s ituations, neonates and tissue subpopulations (e.g., laser cell microdissected biopsy samples). This level of miniaturization translates to minimally invasive yet data-rich molecular diagnostics for early detection of cancer and other diseases with a significant reduction in patient discomfort.