Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
J Chromatogr A. 2012 Oct 26;1261:129-35. doi: 10.1016/j.chroma.2012.08.095. Epub 2012 Sep 1.
The integration of sample preparation methods into microfluidic devices provides automation necessary for achieving complete micro total analysis systems. We have developed a technique that combines on-chip sample enrichment with fluorescence labeling and purification. Polymer monoliths made from butyl methacrylate were fabricated in cyclic olefin copolymer microdevices and used for solid phase extraction. We studied the retention of fluorophores, amino acids and proteins on these columns. The retained samples were subsequently labeled with both Alexa Fluor 488 and Chromeo P503, and unreacted dye was rinsed off the column before sample elution. Additional purification was obtained from the differential retention of proteins and fluorescent labels. A linear relation between the eluted peak areas and concentrations of on-chip labeled heat shock protein 90 samples demonstrated the utility of this method for on-chip quantitation. Our fast and simple method of simultaneously concentrating and labeling samples on-chip is compatible with miniaturization and desirable for automated analysis.
将样品制备方法与微流控装置集成，为实现完整的微全分析系统提供了必要的自动化。我们开发了一种将芯片上样品富集与荧光标记和纯化相结合的技术。采用甲基丙烯酸丁酯制备的聚合物整体柱在环烯烃共聚物微器件中制造，并用于固相萃取。我们研究了这些柱子对荧光团、氨基酸和蛋白质的保留情况。随后，用 Alexa Fluor 488 和 Chromeo P503 对保留的样品进行标记，在洗脱样品之前，用缓冲液将未反应的染料从柱子上冲洗掉。通过蛋白质和荧光标记的差异保留获得了额外的纯化效果。芯片上标记的热休克蛋白 90 样品洗脱峰面积与浓度之间的线性关系证明了该方法用于芯片定量的实用性。我们的快速、简单的方法可以在芯片上同时浓缩和标记样品，与微型化兼容，适用于自动化分析。
