压电(QCM)生物传感器 2009

User-friendly, miniature biosensor flow cell for fragile high fundamental frequency quartz crystal resonators.

Biosensors & bioelectronics Sagmeister BP, Graz IM, Schwödiauer R, Gruber H, Bauer S
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组成图示

User-friendly, miniature biosensor fl... 传感器构成示意图

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传感器类型

压电(QCM)生物传感器

检测对象

磷脂单层(phospholipid monolayer,DOPC/DOPS)、链霉亲和素(streptavidin)、生物素化免疫球蛋白G(biotinylated IgG);样品基质:PBS缓冲液、脂质体水悬浮液、去污剂水溶液。

检测原理

该传感器基于AT-cut石英晶体厚度剪切模式(TSM)的压电共振。表面吸附生物分子引起质量与黏度变化,按Sauerbrey关系Δf = −2f0^2/√(ρqμq)·Δm使共振频率下降;提高基频f0可增强灵敏度。50 MHz HFF晶体表面蒸镀Au/Cr后,1-十八烷硫醇(ODT)形成疏水自组装单层。磷脂脂质体在ODT上铺展形成磷脂单层;若含biotin-cap-DOPE,则提供生物素位点,链霉亲和素结合后进一步结合生物素化IgG,形成蛋白多层。每层质量累积导致频率逐步降低,去污剂改变溶液黏度/密度并去除脂质,引起可逆频率变化。阻抗分析仪每2.5 s读取f0,实现实时监测。

检测灵敏度

原文未报告 LOD、线性范围、灵敏度斜率或相关系数。

效应效果

流动池夹持的50 MHz HFF晶体干态σ=0.67 Hz、Q=9820;液体加载σ=14.70 Hz、Q=1034。PBS基线约50.8549 MHz、σ±10.2 Hz。磷脂单层形成约7 min,Δf≈−560至−700 Hz;40 mM octylglucoside产生约−2 kHz黏度响应,PBS可恢复,PDMS无泄漏。蛋白多层中,含生物素磷脂单层Δf≈−5 kHz,链霉亲和素层平均−6000 Hz,每层生物素化IgG约−4400 Hz。作者称可几秒换片,无需胶接/布线。

传感器的构成

  • 换能器基底:50 MHz高基频石英晶体谐振器(HFF-QCM,AT-cut,TSM),中心薄膜约33.4 μm,提供压电振荡与质量传感
  • 电极层:双面蒸镀70 nm Au/3 nm Cr,Au作为功能化基底并电接触,Cr增强附着
  • 自组装单层:1-十八烷硫醇(ODT)在Au表面形成疏水SAM,用于磷脂单层吸附
  • 磷脂功能层:DOPC/DOPS或DOPC/DOPS/biotin-cap-DOPE脂质体在ODT上形成磷脂单层,提供膜界面或生物素位点
  • 蛋白识别层:链霉亲和素(streptavidin)与生物素化IgG(biotinylated IgG)交替结合,构建蛋白多层
  • 流动池密封与电极:PEEK流道芯、不锈钢基电极、PDMS密封剂、铝结构和顶部平面电极,支撑晶体并密封0.164 μl流室

中文摘要

为将高基频(HFF)石英晶体谐振器用作超灵敏声学生物传感器,作者设计并测试了一种专用石英晶体微天平(QCM)流动池。该流动池可快速、简便地安装和更换小型且易碎的HFF传感器,兼顾机械、热、电和化学要求,并以简单结构解决部分相互矛盾的设计约束。文中讨论了核心设计概念,并简要描述制造过程,重点介绍关键部件的制备。测试采用标准50 MHz HFF谐振器,其表面经1-十八烷硫醇自组装单层功能化。通过两类实验验证其可靠性:一是实时监测磷脂单层在疏水表面的形成以及用去污剂去除;二是通过交替固定链霉亲和素和生物素化免疫球蛋白G,监测蛋白多层的逐步生长。结果表明该流动池能稳定支持HFF-QCM在液相中的生物分子相互作用检测。

英文摘要

For the application of high fundamental frequency (HFF) quartz crystal resonators as ultra sensitive acoustic biosensors, a tailor-made quartz crystal microbalance (QCM) flow cell has been fabricated and tested. The cell permits an equally fast and easy installation and replacement of small and fragile HFF sensors. Usability and simple fabrication are two central features of the HFF-QCM flow cell. Mechanical, thermal, electrical and chemical requirements are considered. The design of the cell combines these, partially contradictory, requirements within a simple device. Central design concepts are discussed and a brief description of the fabrication, with a special focus on the preparation of crucial parts, is provided. For test measurements, the cell was equipped with a standard 50 MHz HFF resonator which had been surface-functionalised with a self-assembled monolayer of 1-octadecanethiol. The reliable performance is demonstrated with two types of experiments: the real time monitoring of phospholipid monolayer formation and its removal with detergent, as well as step-wise growth of a protein multilayer system by an alternating immobilisation of streptavidin and biotinylated immunoglobulin G.

关键词

高基频石英晶体微天平QCM流动池压电生物传感器自组装单层磷脂单层蛋白多层