传感器类型
压电(QCM)生物传感器
检测对象
R,S-1,2,3,4-四氢-1-萘胺(R,S-1-TNA)、R,S-1-(4-甲氧基苯基)乙胺(R,S-4-MPEA)、R,S-1-(3-甲氧基苯基)乙胺(R,S-3-MPEA)、R,S-2-辛醇(R,S-2-OT)、R,S-乳酸甲酯(R,S-MEL);样品基质为气相挥发性蒸气(氮气载气)
检测原理
传感器以自组装固定的GSA或RbSA作为手性识别元件。当气相中对映体蒸气随氮气载气接触传感表面时,对映体分子与白蛋白的手性结合位点发生特异性结合。由于R和S对映体与白蛋白的结合亲和力不同,单位时间内吸附在表面的质量不同。根据Sauerbrey关系,吸附质量增加使QCM共振频率下降,频率变化Δf与吸附质量成正比。分别记录R和S对映体引起的ΔfR和ΔfS,并计算手性判别因子αQCM=ΔfR/ΔfS或ΔfS/ΔfR,从而实现对映体的手性识别与区分。该过程无额外信号放大,主要依赖蛋白手性结合位点的立体选择性。
检测灵敏度
相关系数: R*>0.99(UV/FL结合常数拟合);原文未报告LOD、线性范围、灵敏度斜率
效应效果
该QCM手性传感器对五对对映体表现出良好的手性选择性和可重复性,连续五次测量结果可重复,空白传感器对R/S对映体无手性选择性。GSA传感器的αQCM范围为1.06(R,S-3-MPEA)至1.34(R,S-1-TNA),RbSA传感器为1.11(R,S-3-MPEA和R,S-MEL)至1.28(R,S-1-TNA)。对1-TNA、4-MPEA和3-MPEA,GSA与RbSA的立体选择性偏好一致;对2-OT和MEL则相反,说明识别具有物种依赖性。UV和FL光谱得到的结合常数与手性判别趋势同QCM一致,支持传感机制。作者认为该方法具有质量灵敏度高、分析速度快、成本低、适合实时分析和芯片化应用的潜力。
传感器的构成
- 基底/换能器:AT-cut石英晶体(QCM crystal),5 MHz金镀层,提供压电换能与频率读出
- 自组装修饰层:巯基乙酸(mercaptoacetic acid, MAA)自组装单分子层(SAM),提供羧基终端用于蛋白固定
- 活化处理层:EDC/NHS活化羧基,形成活性酯以共价偶联蛋白
- 识别元件:山羊血清白蛋白(GSA)或兔血清白蛋白(RbSA),作为手性选择剂识别对映体
- 样品引入层:气相脉冲测量系统与氮气载气,使挥发性对映体蒸气接触传感表面
- 信号读出:QCM频率变化(Δf),对映体吸附导致质量增加、频率下降
中文摘要
本研究报道了一种基于石英晶体微天平(QCM)的手性生物传感器,用于五对对映体的手性识别。以山羊血清白蛋白(GSA)和兔血清白蛋白(RbSA)作为手性选择剂,通过自组装单分子层技术将其固定于QCM表面,GSA和RbSA的表面浓度分别为8.8×10^-12 mol cm^-2和1.2×10^-11 mol cm^-2。传感器表现出良好的灵敏度和选择性,但手性识别能力具有明显的物种依赖性。对R,S-1,2,3,4-四氢-1-萘胺、R,S-1-(4-甲氧基苯基)乙胺和R,S-1-(3-甲氧基苯基)乙胺,两种白蛋白传感器的立体选择性结合偏好一致;而对R,S-2-辛醇和R,S-乳酸甲酯,两种传感器呈现相反的手性识别偏好。此外,通过紫外(UV)和荧光(FL)光谱进一步研究了白蛋白与对映体的相互作用,UV/FL结果与QCM手性识别趋势一致。
英文摘要
Quartz crystal microbalance (QCM) biosensor was used for the chiral recognition of five pairs of enantiomers by using goat serum albumin (GSA) and rabbit serum albumin (RbSA) as chiral selectors. Serum albumin (SA) was immobilized on the QCM through the self-assembled monolayer technique, and the surface concentration of GSA and RbSA were 8.8 × 10(-12) mol cm(-2) and 1.2 × 10(-11) mol cm(-2) , respectively. The QCM biosensors showed excellent sensitivity and selectivity. Meanwhile, the chiral recognition of SA sensors was quite species dependent. There were differences between GSA and RbSA sensors in the ability and the preference of chiral recognition. To R,S-1,2,3,4-tetrahydro-1-naphthylamine (R,S-1-TNA), R,S-1-(4-methoxyphenyl)ethylamine (R,S-4-MPEA), and R,S-1-(3-methoxyphenyl)ethylamine (R,S-3-MPEA), the preference of the stereoselective SA-drug binding of the two kinds of SA sensors were consistent. However, to R,S-2-octanol (R, S-2-OT) and R,S-methyl lactate (R,S-MEL), the two kinds of SA sensors had opposite chiral recognition preference. Moreover, the interactions of SA and the five pairs of enantiomers have been further investigated through ultraviolet (UV) and fluorescent (FL) spectra. The UV/FL results were in accordance with the consequence of QCM.