Fan Z Hugh
Department of Mechanical and Aerospace Engineering, University of Florida, Gainesville, FL 32611, USA.
Methods Mol Biol. 2009;544:43-52. doi: 10.1007/978-1-59745-483-4_4.
Plastic microfluidic devices are fabricated with an array of pseudo-valves for two-dimensional (2D) protein separation. The devices are made by compression molding; the mold is created by electroplating on a glass master fabricated by photolithography. Each device consists of one channel for isoelectric focusing (IEF) and multiple parallel channels for polyacrylamide gel electrophoresis (PAGE). The IEF channel (first dimension) is orthogonal to the PAGE channels (second dimension). Microfluidic pseudo-valves are created at the intersections of orthogonal channels by photodefinable, in situ gel polymerization. These valves enable the introduction of two types of separation media into orthogonal channels for performing 2D protein separation in the device. The presence of the pseudo-valves prevents one separation medium from being contaminated by the other medium, although proteins are allowed to transfer from the first to the second dimension under an electric field. Two-dimensional protein separation is achieved in less than 10 min, an improvement of two orders of magnitude compared with the conventional 2D gel electrophoresis using an IEF strip and a PAGE slab.
塑料微流控装置通过一系列伪阀制造而成，用于二维（2D）蛋白质分离。这些装置通过压缩成型制成；模具通过在光刻制造的玻璃母版上电镀产生。每个装置由一个用于等电聚焦（IEF）的通道和多个用于聚丙烯酰胺凝胶电泳（PAGE）的平行通道组成。IEF通道（第一维）与PAGE通道（第二维）正交。通过光可定义的原位凝胶聚合在正交通道的交叉点处形成微流控伪阀。这些阀能够将两种类型的分离介质引入正交通道，以便在装置中进行二维蛋白质分离。尽管在电场作用下蛋白质可以从第一维转移到第二维，但伪阀的存在可防止一种分离介质被另一种介质污染。二维蛋白质分离可在不到10分钟内完成，与使用IEF条带和PAGE平板的传统二维凝胶电泳相比，提高了两个数量级。
