Goyon Alexandre, Francois Yannis Nicolas, Colas Olivier, Beck Alain, Veuthey Jean Luc, Guillarme Davy
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Geneva, Switzerland.
Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS) UMR 7140 (Unistra-CNRS), Université de Strasbourg, Strasbourg, France.
Electrophoresis. 2018 Aug;39(16):2083-2090. doi: 10.1002/elps.201800131. Epub 2018 Jun 4.
The determination of mAb critical quality attributes (CQA) is crucial for their successful application in health diseases. A generic CZE method was developed for the high-resolution separation of various mAb charge variants, which are often recognized as important CQA. A dynamic coating of the capillary was obtained with polyethylene oxide (PEO), whereas Bis-Tris allowed the analysis of mAbs under native conditions at pH 7.0. The effect of PEO and Bis-Tris concentrations, as well as the nature of the acidic counter ion on the method performance was systematically studied. The %RSD on migration times was below 5% on three different CE instruments using the optimized method. Additional charge variants (in particular acidic variants) were resolved for 10 out of 17 mAbs compared to a reference CZE approach involving the use of ε-amino-caproic acid (EACA), triethylenetetramine (TETA), and hydroxypropylmethyl cellulose (HPMC). The amount of basic and acidic charge variants of 17 Food and Drug Administration (FDA) approved mAbs covering a wide range of physico-chemical properties, e.g., pI between 8.0 and 9.4 and different hydrophobicity, were mainly comprised between 5-15% and 15-30%, respectively. It is noteworthy that applications for the quality control in hospitals as well as for the combination of the immune checkpoint inhibitors nivolumab and ipilimumab were presented.
单克隆抗体关键质量属性（CQA）的确定对于其在健康疾病中的成功应用至关重要。开发了一种通用的毛细管区带电泳（CZE）方法，用于高分辨率分离各种单克隆抗体电荷变体，这些变体通常被认为是重要的CQA。用聚环氧乙烷（PEO）对毛细管进行动态涂层，而Bis-Tris允许在pH 7.0的天然条件下分析单克隆抗体。系统研究了PEO和Bis-Tris浓度以及酸性抗衡离子的性质对方法性能的影响。使用优化方法，在三种不同的毛细管电泳仪器上，迁移时间的相对标准偏差（%RSD）低于5%。与涉及使用ε-氨基己酸（EACA）、三乙烯四胺（TETA）和羟丙基甲基纤维素（HPMC）的参考CZE方法相比，17种单克隆抗体中有10种解析出了额外的电荷变体（特别是酸性变体）。17种美国食品药品监督管理局（FDA）批准的单克隆抗体，涵盖了广泛的物理化学性质，例如，pI在8.0至9.4之间且疏水性不同，其碱性和酸性电荷变体的含量分别主要在5-15%和15-30%之间。值得注意的是，还介绍了该方法在医院质量控制以及免疫检查点抑制剂纳武单抗和伊匹单抗联合应用方面的情况。
