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使用紫外光接枝法在由聚丙烯酰胺涂覆的环状烯烃共聚物微流体通道中进行等电聚焦。

Isoelectric focusing in cyclic olefin copolymer microfluidic channels coated by polyacrylamide using a UV photografting method.

作者信息

Li Chen, Yang Yanou, Craighead Harold G, Lee Kelvin H

机构信息

School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853, USA.

出版信息

Electrophoresis. 2005 May;26(9):1800-6. doi: 10.1002/elps.200410309.

Abstract

As an alternative material to glass or silicon, microfluidic devices made from a cyclic olefin copolymer (COC) were fabricated. This material is of interest because of the relative ease of fabrication, low costs, and solvent resistance. However, as a result of the strong hydrophobic interactions normally present, COC surfaces are not suitable for protein separations. To reduce the protein adsorption and make COC suitable for protein separations, UV-initiated grafting of polyacrylamide was used to coat the surface of COC devices. The change in surface properties caused by different graft times was studied. The surface hydrophilicity and electroosmotic mobility were characterized by contact angle and electroosmosis measurements. Isoelectric focusing was performed to test protein separations in polyacrylamide-coated COC microchannels. A single protein, carbonic anhydrase, was used to analyze the focusing effects and peak capacities in uncoated and polyacrylamide-coated COC devices. Peak capacities ranging from 75 to 190 were achieved with a polyacrylamide-coated surface. A mixture of two proteins, conalbumin labeled with Alexa Fluor 488 and beta-lactoglobulin A labeled with Alexa Fluor 546, was used to test protein separations. Linear and rapid separation of proteins was achieved in the polyacrylamide-coated COC microfluidic device.

摘要

作为玻璃或硅的替代材料,制备了由环烯烃共聚物(COC)制成的微流控装置。这种材料因其相对易于制造、成本低和耐溶剂性而备受关注。然而,由于通常存在的强疏水相互作用,COC表面不适用于蛋白质分离。为了减少蛋白质吸附并使COC适用于蛋白质分离,采用紫外线引发的聚丙烯酰胺接枝来涂覆COC装置的表面。研究了不同接枝时间引起的表面性质变化。通过接触角和电渗测量对表面亲水性和电渗迁移率进行了表征。进行等电聚焦以测试聚丙烯酰胺涂层的COC微通道中的蛋白质分离。使用单一蛋白质碳酸酐酶来分析未涂覆和聚丙烯酰胺涂覆的COC装置中的聚焦效果和峰容量。聚丙烯酰胺涂层表面的峰容量范围为75至190。使用两种蛋白质的混合物,即标记有Alexa Fluor 488的伴清蛋白和标记有Alexa Fluor 546的β-乳球蛋白A,来测试蛋白质分离。在聚丙烯酰胺涂层的COC微流控装置中实现了蛋白质的线性快速分离。

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