Institute of Engineering Materials and Design, Faculty of Mechanical Engineering, University of Maribor, Maribor, Slovenia.
Electrophoresis. 2010 Mar;31(6):1097-100. doi: 10.1002/elps.200900525.
Protein-protein conjugates formed by carbodiimide crosslinking reactions have been analyzed for the first time using CE. Lysozyme and BSA were chosen as model proteins to study the efficacy of N-(3-dimethylaminopropyl)-N-ethylcarbodiimide and N-hydroxysuccinimide as crosslinkers. Detection of the molecular mass increase was checked by SDS-PAGE. Commercially available, PVA-coated capillaries showed appropriate selection, while phospho-deactivated and dynamic PVA-coated capillaries did not give suitable resolution. CE was found to be an efficient tool to characterize homo- (lysozyme-lysozyme) and hetero- (lysozyme-BSA) protein coupling by suitable variations of electrophoretic mobilities.
首次使用 CE 分析了通过碳二亚胺交联反应形成的蛋白质-蛋白质缀合物。选择溶菌酶和 BSA 作为模型蛋白，研究 N-(3-二甲基氨基丙基)-N-乙基碳二亚胺和 N-羟基琥珀酰亚胺作为交联剂的效果。通过 SDS-PAGE 检查了分子质量增加的检测。商业上可用的 PVA 涂层毛细管显示出适当的选择，而磷酸化失活和动态 PVA 涂层毛细管则不能给出合适的分辨率。通过适当改变电泳迁移率，CE 被发现是一种有效的工具，可用于表征同型（溶菌酶-溶菌酶）和异型（溶菌酶-BSA）蛋白质偶联。
