通过毛细管电泳电喷雾电离质谱法（CE-ESI-MS）筛选小完整蛋白质。

Screening of Small Intact Proteins by Capillary Electrophoresis Electrospray Ionization-Mass Spectrometry (CE-ESI-MS).

作者信息

Neuberger Sabine, Rafai Angelina, Neusüß Christian

机构信息

Department of Chemistry, Aalen University, Beethovenstraβe 1, Aalen, D-73430, Germany.

Solvias AG, Kaiseraugst, Switzerland.

出版信息

Methods Mol Biol. 2016;1466:43-56. doi: 10.1007/978-1-4939-4014-1_4.

DOI:10.1007/978-1-4939-4014-1_4

PMID:27473480

Abstract

Capillary electrophoresis (CE) has been shown to be a suitable separation technique for complex samples. Combined with electrospray ionization-mass spectrometry (ESI-MS), it is a powerful tool offering the opportunity of high selectivity and sensitivity combined with the possibility to identify and characterize intact proteins. In this protocol, we demonstrate a screening method for intact proteins based on capillary zone electrophoresis (CZE) separation coupled with online mass spectrometric detection. In order to avoid protein-wall interactions, a neutral coated capillary is used to create a universal method for proteins with both low and high electrophoretic mobilities. In addition, we show the successful validation and application of this screening method for a set of eight standard proteins and the glycoprotein erythropoietin.

摘要

毛细管电泳（CE）已被证明是一种适用于复杂样品的分离技术。与电喷雾电离质谱（ESI-MS）相结合，它是一种强大的工具，提供了高选择性和灵敏度的机会，同时具备识别和表征完整蛋白质的可能性。在本方案中，我们展示了一种基于毛细管区带电泳（CZE）分离并结合在线质谱检测的完整蛋白质筛选方法。为避免蛋白质与管壁相互作用，使用中性涂层毛细管创建一种适用于电泳迁移率低和高的蛋白质的通用方法。此外，我们展示了该筛选方法对一组八种标准蛋白质和糖蛋白促红细胞生成素的成功验证及应用。