Shim Ji Wook, Gu Li Qun
Department of Biological Engineering and Dalton Cardiovascular Research Center, University of Missouri, Columbia, Missouri 65211, USA.
Anal Chem. 2007 Mar 15;79(6):2207-13. doi: 10.1021/ac0614285. Epub 2007 Feb 9.
Engineered protein channels have many potential applications in biosensing at the single-molecule level. A future generation of biosensor could be an array of target-specific ion channels, where each protein pore acts as a sensor element. An important step toward this goal is to create a portable, durable, single-protein channel-integrated chip device. Here we report a versatile, modular chip that contains a single-ion channel for single-molecular biosensing. The core of the device is a long-lived lipid membrane that has been sandwiched between two air-insulated agarose layers which gel in situ. A single-protein pore embedded in the membrane serves as the sensor element. The modular device is highly portable, allowing a single-ion channel to continuously function following detachment of the chip from the instrument and independent transportation of the device. The chip also exhibits high durability, which is evidenced from long-duration continuous observation of single-channel dynamics. Once engineered protein pores are installed, the chip becomes a robust stochastic sensor for real-time targeting such as detection of the second messenger IP3. This pluggable biochip could be incorporated with many applicable devices, such as a microfluidic system, and be made into a microarray for both biomedical detection and membrane protein research.
工程化蛋白质通道在单分子水平的生物传感中有许多潜在应用。下一代生物传感器可能是由目标特异性离子通道组成的阵列,其中每个蛋白质孔充当一个传感元件。朝着这个目标迈出的重要一步是创建一种便携式、耐用的、集成单蛋白质通道的芯片装置。在此,我们报告一种通用的模块化芯片,其包含用于单分子生物传感的单离子通道。该装置的核心是一个长寿命脂质膜,它夹在两个原位凝胶化的空气绝缘琼脂糖层之间。嵌入膜中的单个蛋白质孔用作传感元件。该模块化装置具有高度便携性,芯片从仪器上分离并独立运输后,单离子通道仍能持续发挥作用。该芯片还具有高耐用性,这在对单通道动力学的长时间连续观察中得到了证明。一旦安装了工程化蛋白质孔,该芯片就成为一种强大的随机传感器,可用于实时靶向检测,如检测第二信使IP3。这种可插拔生物芯片可以与许多适用装置(如微流体系统)结合,并制成用于生物医学检测和膜蛋白研究的微阵列。