Yao S, Anex D S, Caldwell W B, Arnold D W, Smith K B, Schultz P G
Department of Chemistry, Scripps Research Institute, La Jolla, CA 92037, USA.
Proc Natl Acad Sci U S A. 1999 May 11;96(10):5372-7. doi: 10.1073/pnas.96.10.5372.
Analysis of variations in the concentrations or structures of biomolecules (e.g., mRNAs, proteins, peptides, natural products) that occur either naturally or in response to environmental or genetic perturbations can provide important insight into complex biological processes. Many biological samples are mixtures that require a separation step before quantitation of variations in the individual components. Two-dimensional denaturing gel electrophoresis has been used very effectively to separate complex mixtures of proteins, but it is time consuming and requires considerable amounts of sample. Microchannel-based separations have proven very effective in rapidly separating small amounts of nucleic acids; more recently, isoelectric focusing of proteins also has been adapted to the microchannel format. Here, we describe microchannel-based SDS capillary gel electrophoresis of proteins and demonstrate the speed and high resolution it provides. This development is an important step toward the miniaturization and integration of multidimensional and array separation methods for complex protein mixtures.
分析天然存在或响应环境或基因扰动而发生的生物分子(如mRNA、蛋白质、肽、天然产物)浓度或结构的变化,可为复杂生物过程提供重要见解。许多生物样品是混合物,在定量单个成分的变化之前需要进行分离步骤。二维变性凝胶电泳已非常有效地用于分离复杂的蛋白质混合物,但它耗时且需要大量样品。基于微通道的分离已被证明在快速分离少量核酸方面非常有效;最近,蛋白质的等电聚焦也已适用于微通道形式。在此,我们描述了基于微通道的蛋白质SDS毛细管凝胶电泳,并展示了其提供的速度和高分辨率。这一进展是朝着复杂蛋白质混合物的多维和阵列分离方法的小型化和集成迈出的重要一步。