Microcantilever-Based Label-Free Characterization of Temperature-Dependent Biomolecular Affinity Binding.
Microcantilever-Based Label-Free Characterization of Temperature-Dependent Biomolecular Affinity Binding.
复制标题
DOI：
10.1016/j.snb.2012.02.045
复制
发表时间：
2013-01-01
影响因子：
8.4
通讯作者：
Lin, Qiao
中科院分区：
文献类型：
--
作者：
Wang, Bin;Huang, Fengliang;ThaiHuu Nguyen;Xu, Yong;Lin, Qiao
本文介绍了使用微悬臂装置对固体表面上温度依赖性生物分子亲和结合的无标记表征。该装置由一个聚对二甲苯悬臂梁组成，其一侧涂有一层金膜，并与分子功能化，作为目标分析物的亲和受体。悬臂位于聚二甲基硅氧烷（PDMS）微流控室中，该微流控室与底层衬底上的透明氧化铟锡（ITO）电阻式温度传感器集成在一起。ITO传感器允许实时测量腔室温度，以及无阻碍的光学通道，用于基于反射的悬臂挠度光学检测。为了测试靶和受体之间的温度依赖性结合，将腔室的温度保持在恒定的设定值，同时将未标记的分析物分子溶液连续注入腔室。测量的悬臂挠度用于确定目标-受体结合特性。我们展示了血小板衍生生长因子（PDGF）蛋白与适体受体的温度依赖性结合动力学的无标记表征。得到了亲和结合特性，包括缔合和解离速率常数以及平衡解离常数，并显示出对温度的显著依赖。
This paper presents label-free characterization of temperature-dependent biomolecular affinity binding on solid surfaces using a microcantilever-based device. The device consists of a Parylene cantilever one side of which is coated with a gold film and functionalized with molecules as an affinity receptor to a target analyte. The cantilever is located in a poly(dimethylsiloxane) (PDMS) microfluidic chamber that is integrated with a transparent indium tin oxide (ITO) resistive temperature sensor on the underlying substrate. The ITO sensor allows for real-time measurements of the chamber temperature, as well as unobstructed optical access for reflection-based optical detection of the cantilever deflection. To test the temperature-dependent binding between the target and receptor, the temperature of the chamber is maintained at a constant setpoint, while a solution of unlabeled analyte molecules is continuously infused through the chamber. The measured cantilever deflection is used to determine the target-receptor binding characteristics. We demonstrate label-free characterization of temperature-dependent binding kinetics of the platelet-derived growth factor (PDGF) protein with an aptamer receptor. Affinity binding properties including the association and dissociation rate constants as well as equilibrium dissociation constant are obtained, and shown to exhibit significant dependencies on temperature.
登录
查看更多内容
影响因子：
7.4
作者：
Savran, CA;Knudsen, SM;Manalis, SR
通讯作者：
Manalis, SR
影响因子：
2.8
作者：
Nguyen, ThaiHuu;Pei, Renjun;Lin, Qiao
通讯作者：
Lin, Qiao
影响因子：
2.9
作者：
Dam, TK;Roy, R;Brewer, CF
通讯作者：
Brewer, CF
使用微流体悬臂阵列测量蛋白酶活性和抑制的定量和无标记技术。
影响因子：
10.8
作者：
Raorane DA;Lim MD;Chen FF;Craik CS;Majumdar A
通讯作者：
Majumdar A
影响因子：
2.8
作者：
De Crescenzo, Gregory;Boucher, Cyril;Jolicoeur, Mario
通讯作者：
Jolicoeur, Mario