Astorga-Wells Juan, Vollmer Susanne, Tryggvason Sam, Bergman Tomas, Jörnvall Hans
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Anal Chem. 2005 Nov 15;77(22):7131-6. doi: 10.1021/ac050931z.
A separation method based on electroimmobilization and sequential release of captured molecules is reported. A microfluidic electrocapture device is utilized to immobilize peptides in a microflow stream. After capture, the electric field is decreased in a stepwise manner, causing sequential release of the captured peptides according to their electrophoretic mobility. Tryptic peptides were separated and analyzed by matrix-assisted laser desorption/ionization mass spectrometry. The separation power was high enough to increase the ionization yield of several peptides not seen in the unprocessed sample. In addition to separation, simultaneous sample cleanup was demonstrated for peptides obtained by shotgun tryptic digestion of membrane protein extracts.
报道了一种基于电固定和捕获分子顺序释放的分离方法。利用微流控电捕获装置在微流中固定肽段。捕获后，电场逐步降低，使捕获的肽段根据其电泳迁移率顺序释放。用基质辅助激光解吸/电离质谱对胰蛋白酶肽段进行分离和分析。分离能力足够高，可提高未处理样品中未见的几种肽段的电离产率。除分离外，还证明了对通过膜蛋白提取物的鸟枪法胰蛋白酶消化获得的肽段可同时进行样品净化。
