Brown Abby, Morales Christina, Gomez Frank A
Department of Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, CA 90032-8202, USA.
Talanta. 2008 Jan 15;74(4):605-12. doi: 10.1016/j.talanta.2007.06.043. Epub 2007 Jul 7.
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.
在本文中，我们描述了一种微流控/毛细管电泳（CE）技术的发展，该技术采用部分填充亲和毛细管电泳（PFACE），以碳酸酐酶B（CAB，EC 4.2.1.1）和东方链霉菌的万古霉素作为模型系统来估计配体与受体的结合常数。使用多层软光刻（MSL）制造了一个由流体通道和控制通道组成的微流控装置（MD），并配备了一个外部毛细管柱。多个流动通道允许将含有配体、受体和非相互作用标准物的样品区带操纵到MD中，随后进入毛细管柱。电泳时，样品组分迁移到配体区带，在那里建立平衡。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上对少量溶液的操纵最大限度地减少了耗时的移液步骤的需求。
