Through-a-chip partial filling affinity capillary electrophoresis for estimating binding constants of ligands to receptors.
Through-a-chip partial filling affinity capillary electrophoresis for estimating binding constants of ligands to receptors.
复制标题
通过芯片部分填充亲和毛细管电泳，用于估计配体与受体的结合常数。
DOI：
10.1016/j.talanta.2007.06.043
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发表时间：
2008
期刊：
影响因子：
6.1
通讯作者：
Gomez,FrankA
中科院分区：
文献类型：
--
作者：
Brown,Abby;Morales,Christina;Gomez,FrankA
本文采用部分填充亲和毛细管电泳法(PFACE)，以碳酸氢酶B(CAB，EC 4.2.1.1)和东方链霉菌的万古霉素为模型体系，建立了一种微流控/毛细管电泳法(CE)测定配体与受体的结合常数。利用多层软光刻(MSL)技术制作了由流体和控制通道组成的微流控器件(MD)，并在其上安装了外置毛细管柱。多个流动通道允许将含有受体和非相互作用标准的配位体和样品的区域操纵到MD中，然后进入毛细管柱。在电泳法上，样品组分迁移到建立平衡的配位体区域。在分析中使用受体迁移时间的变化来获得结合相互作用的值。在MD上操作小体积的溶液可以最大限度地减少耗时的移液步骤。
In this paper, we describe the development of a microfluidic/capillary electrophoresis (CE) technique employing partial filling affinity capillary electrophoresis (PFACE) to estimate binding constants of ligands to receptors using as model systems carbonic anhydrase B (CAB, EC 4.2.1.1) and vancomycin from Streptomyces orientalis. Using multilayer soft lithography (MSL), a microfluidic device (MD) consisting of fluid and control channels is fabricated and fitted with an external capillary column. Multiple flow channels allows for manipulation of a zone of ligand and sample containing receptor and non-interacting standards into the MD and subsequently into the capillary column. Upon electrophoresis the sample components migrate into the zone of ligand where equilibrium is established. Changes in migration time of the receptor are used in the analysis to obtain a value for the binding interaction. The manipulation of small volumes of solution on the MD minimizes the need of time-consuming pipetting steps.