Suba Dávid, Urbányi Zoltán, Salgó András
Chemical Works of Gedeon Richter Plc, 1103 Budapest, Gyömrői út 19-21, Hungary, Hungary.
Chemical Works of Gedeon Richter Plc, 1103 Budapest, Gyömrői út 19-21, Hungary, Hungary.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Oct 1;1032:224-229. doi: 10.1016/j.jchromb.2016.07.026. Epub 2016 Jul 25.
Capillary electrophoresis techniques are widely used in the analytical biotechnology. Different electrophoretic techniques are very adequate tools to monitor size-and charge heterogenities of protein drugs. Method descriptions and development studies of capillary zone electrophoresis (CZE) have been described in literature. Most of them are performed based on the classical one-factor-at-time (OFAT) approach. In this study a very simple method development approach is described for capillary zone electrophoresis: a "two-phase-four-step" approach is introduced which allows a rapid, iterative method development process and can be a good platform for CZE method. In every step the current analytical target profile and an appropriate control strategy were established to monitor the current stage of development. A very good platform was established to investigate intact and digested protein samples. Commercially available monoclonal antibody was chosen as model protein for the method development study. The CZE method was qualificated after the development process and the results were presented. The analytical system stability was represented by the calculated RSD% value of area percentage and migration time of the selected peaks (<0.8% and <5%) during the intermediate precision investigation.
毛细管电泳技术在分析生物技术中被广泛应用。不同的电泳技术是监测蛋白质药物大小和电荷异质性的非常合适的工具。毛细管区带电泳（CZE）的方法描述和开发研究已在文献中有所记载。其中大多数是基于经典的一次一因素（OFAT）方法进行的。在本研究中，描述了一种用于毛细管区带电泳的非常简单的方法开发方法：引入了一种“两阶段四步骤”方法，该方法允许进行快速、迭代的方法开发过程，并且可以成为CZE方法的良好平台。在每个步骤中，都建立了当前的分析目标概况和适当的控制策略，以监测当前的开发阶段。建立了一个很好的平台来研究完整和消化后的蛋白质样品。选择市售单克隆抗体作为方法开发研究的模型蛋白。在开发过程之后对CZE方法进行了验证，并给出了结果。在中间精密度研究期间，通过计算所选峰的面积百分比和迁移时间的RSD%值（<0.8%和<5%）来表示分析系统的稳定性。
