BioMarker Analysis from Single Cells using Nanobiosensor

描述(申请人提供)：这项第一阶段可行性研究的目的是设计、建造和评估一种能够从几个细胞以及最终从单个细胞中检测ng蛋白生物标记物的超微型传感器设备。该装置被称为纳米生物传感器，由印刷在微型制造硅结构上的一系列超微型蛋白质捕获(抗体)结构域组成。纳米生物传感器结合了纳米技术、细胞I和分子生物学的概念、方法和仪器，创造了一条通向单细胞蛋白质组分析的途径。该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S将测试直接带到样品来源(在这种情况下，是一个细胞)，消除了将极少量的样品提取物运送到远程测试地点的困难。相反的研究是由申请机构开发的用于构建生物芯片的纳米阵列技术，其空间地址为超微(1-20 mm)到亚微米(250 Nm)(对应于传统微射线测试所需表面积的1/1000-1/10,000)，并且只需要极小的样品体积(亚微升到皮升)。这种纳米生物传感器将能够分析以前只能通过核酸的聚合酶链式反应扩增来分析的样品的蛋白质图谱。建议的纳米生物传感器开启了对微小蛋白质样本的快速和间接分析的大门，例如在法医S治疗、新生儿和组织亚群中发现的那些样本(例如激光细胞显微解剖活组织样本)。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.