Kelly J F, Locke S J, Ramaley L, Thibault P
Chemistry Department, Dalhousie University, Halifax, Nova Scotia, Canada.
J Chromatogr A. 1996 Jan 12;720(1-2):409-27. doi: 10.1016/0021-9673(94)01197-4.
A capillary electrophoresis (CE) method using acidic buffers and capillaries coated with Polybrene, a cationic polymer has been developed for the separation of glycoproteins and glycopeptides. Electrophoretic conditions have been optimized to provide resolution of individual glycoforms observed for different glycoprotein preparations. These conditions were found to be entirely compatible with the operation of electrospray mass spectrometry (ESMS), which facilitated the assignments of possible carbohydrate compositions of glycopeptides arising from digests of glycoproteins. By using operating conditions enhanced the formation of oxonium fragment ions prior to mass spectral analysis, selective identification of glycopeptides was achieved for complex samples such as those from proteolytic digests or chemical cleavages. Examples of applications are presented for ribonuclease B, ovalbumin, horseradish peroxidase, and a lectin from Erithrina corallodendron using both CE-ESMS and CE with ultraviolet detection (CE-UV).
一种使用酸性缓冲液和涂有聚凝胺（一种阳离子聚合物）的毛细管的毛细管电泳（CE）方法已被开发用于糖蛋白和糖肽的分离。电泳条件已得到优化，以实现对不同糖蛋白制剂中观察到的各个糖型的分离。发现这些条件与电喷雾质谱（ESMS）的操作完全兼容，这有助于确定糖蛋白消化产生的糖肽可能的碳水化合物组成。通过使用在质谱分析之前增强氧鎓碎片离子形成的操作条件，对于诸如来自蛋白水解消化或化学裂解的复杂样品实现了糖肽的选择性鉴定。使用CE-ESMS和带紫外检测的CE（CE-UV）展示了核糖核酸酶B、卵清蛋白、辣根过氧化物酶和来自珊瑚刺桐的凝集素的应用实例。
