Department of Chemistry, York University, Toronto, Ontario, Canada.
Anal Chem. 2011 Aug 15;83(16):6330-5. doi: 10.1021/ac201242r. Epub 2011 Jul 19.
Methods of kinetic capillary electrophoresis (KCE) facilitate highly efficient selection of DNA aptamers for protein targets. The inability to detect native proteins at low concentrations in capillary electrophoresis creates, however, a significant obstacle for many important protein targets. Here we suggest that protein labeling with new Chromeo dyes can help to overcome this obstacle. By labeling a number of proteins with Chromeo P503, we show that the labeling procedure enables accurate detection of proteins in CE without significantly affecting their electrophoretic mobility or their ability to bind DNA. Moreover, Chromeo P503 does not appear to label the amino-groups of buffer components to a significant extent, making the labeling procedure compatible with a large number of selection and run buffers. Fluorescent labeling of protein targets with Chromeo dyes empowers selection of aptamers by KCE methods and promises to increase the rate at which aptamers for new targets are being developed and introduced in various applications.
动力学毛细管电泳（KCE）方法可高效选择适用于蛋白质靶标的 DNA 适体。然而，在毛细管电泳中无法检测到低浓度的天然蛋白质，这对许多重要的蛋白质靶标构成了重大障碍。在这里，我们提出蛋白质用新型 Chromeo 染料进行标记可以帮助克服这一障碍。通过用 Chromeo P503 标记许多蛋白质，我们表明标记程序可在不显著影响其电泳迁移率或与 DNA 结合能力的情况下，在 CE 中准确检测蛋白质。此外，Chromeo P503 似乎不会显著标记缓冲成分的氨基基团，从而使标记程序与许多选择和运行缓冲液兼容。用 Chromeo 染料对蛋白质靶标进行荧光标记可通过 KCE 方法增强适体的选择，并有望提高针对新靶标的适体的开发速度，并在各种应用中引入。
