Espinosa-de la Garza Carlos Eduardo, Salazar-Flores Rodolfo Daniel, Pérez Néstor Octavio, Flores-Ortiz Luis Francisco, Medina-Rivero Emilio
Unidad de Investigación y Desarrollo, Probiomed S.A. de C.V.
Unidad de Investigación y Desarrollo, Probiomed S.A. de C.V.;
J Vis Exp. 2017 Jan 16(119):55082. doi: 10.3791/55082.
Biotherapeutic proteins, such as monoclonal antibodies (mAbs), are feasible alternatives for the treatment of chronic-degenerative diseases. The biological activity of these proteins depends on their physicochemical properties. The use of high-performance techniques like chromatography and capillary electrophoresis has been described for the analysis of physicochemical heterogeneity of mAbs. Nowadays, capillary zone electrophoresis (CZE) technique constitutes one of the most resolutive and sensitive assays for the analysis of biomolecules. Besides, the electro-driven separation in CZE is governed by extensive properties of matter and offers the advantage of analyzing proteins close to their native state. However, the successful implementation of this technique for routine analysis depends on the skills of the analyst at the critical steps during sample and system preparation. The purpose of this tutorial is to detail the steps to succeed in the CZE analysis of mAbs. Further, this protocol can be used for the development and improvement of skills of the personnel involved in protein analytical chemistry laboratories.
生物治疗蛋白，如单克隆抗体（mAbs），是治疗慢性退行性疾病的可行替代方案。这些蛋白质的生物活性取决于其物理化学性质。已描述了使用色谱法和毛细管电泳等高性能技术来分析单克隆抗体的物理化学异质性。如今，毛细管区带电泳（CZE）技术是分析生物分子最具分辨率和灵敏度的方法之一。此外，CZE中的电驱动分离受物质的广泛性质支配，并具有分析接近其天然状态的蛋白质的优势。然而，该技术在常规分析中的成功实施取决于分析人员在样品和系统制备关键步骤中的技能。本教程的目的是详细介绍在单克隆抗体的CZE分析中取得成功的步骤。此外，该方案可用于蛋白质分析化学实验室相关人员技能的培养和提升。
