Nanometer sized sensors with biological transducers

描述（由申请人提供）：在过去的二十年里，在复杂生物环境的分析方面取得了重大进展。利用指示剂染料或蛋白质的荧光成像已经发挥了重要作用，但也受到显著的限制，包括毒性、化学稳定性和系统的扰动。虽然对于荧光指示剂或离子载体可用的各种离子和小分子非常有用，但由于缺乏合适的指示剂或缺乏固有的光学或电化学活性，大类化学和生物学相关的分析物无法检测。膜受体、离子通道和分子转运蛋白在化学传感应用中的使用呈现出优于传统报告物的几个潜在优点，因为许多这样的膜蛋白对基于小分子和/或蛋白质的配体具有高度选择性，并且通常表现出可转化为传感器的高灵敏度的高亲和力结合。在这里，我们提出了一类新的纳米尺寸，仿生化学传感器与嵌入膜蛋白的细胞内，在体内和环境标记，跟踪和传感应用。为了实现这样的传感器平台，我们将从一系列可以化学交联的脂质中创建磷脂囊泡，并且我们将用重构的膜蛋白使囊泡功能化。我们的传感器平台的关键特征是：a）由可聚合脂质和脂质复合物的自组装形成的磷脂膜; B）并入的生物信号转导元件，例如受体; c）用于启动细胞摄取和传感器定位的外部膜元件;和d）用于产生光学信号的指示剂或报告元件。这种模块化的传感器几何结构将提供一个通用平台，可以用来容易地设计各种各样的传感器。

DESCRIPTION (provided by applicant): The past two decades have witnessed significant advances in the analysis of complex biological environments. Fluorescence imaging utilizing indicator dyes or proteins has played an important role but also suffers from significant limitations including toxicity, chemical stability and perturbation of the system. Though quite useful for a variety of ions and small molecules where fluorescent indicators or ionophores are available, large categories of chemically and biologically relevant analytes elude detection due to a lack of suitable indicators or lack of intrinsic optical or electrochemical activity. The use of membrane receptors, ion channels and molecular transporters in chemical sensing applications presents several potential advantages over traditional reporters as many such membrane proteins are highly selective to small molecule and/or protein based ligands and typically demonstrate high affinity binding that may translate to high sensitivity of the sensor. Here we propose a new class of nanometer-sized, biomimetic chemical sensors with embedded membrane proteins for intracellular, in vivo and environmental labeling, tracing and sensing applications. To realize such a sensor platform, we will create phospholipid vesicles from a series of lipids that can be chemically crosslinked and we will functionalize the vesicles with reconstituted membrane proteins. The key features of our sensor platform are: a) a phospholipid membrane formed from self assembly of polymerizable lipids and lipid composites; b) incorporated biological signal transduction elements, e.g. receptors; c) external membrane elements that serve to initiate cellular uptake and localization of the sensor; and d) indicator or reporter elements that serve to generate an optical signal. This modular sensor geometry will provide a general platform that can be utilized to readily design a wide variety of sensors.