Fekete Szabolcs, Beck Alain, Guillarme Davy
School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Boulevard d'Yvoy 20, 1211 Geneva 4, Switzerland.
Center of Immunology Pierre Fabre, 5 Avenue Napoléon III, BP 60497, 74160 Saint-Julien-en-Genevois, France(1).
J Pharm Biomed Anal. 2015;111:169-76. doi: 10.1016/j.jpba.2015.03.041. Epub 2015 Apr 7.
Cation exchange chromatography (CEX) is a well established strategy for the characterization of monoclonal antibodies (mAbs). The optimization of mobile phase conditions is well described in the literature, but there is a lack of information about CEX stationary phases for the analysis of therapeutic proteins. The aim of this study was to compare five state-of-the-art CEX stationary phases based on the retention, selectivity and resolving power achieved in pH- and salt-gradient modes, with various therapeutic mAbs and their variants. The Sepax Antibodix WCX-NP3, Thermo MAbPac SCX-10 RS, YMC BioPro SP-F, Waters Protein-Pak Hi Res SP and Agilent Bio mAb NP1.7 SS were considered in this study. In terms of retention, the YMC Bio Pro SP-F material was the less retentive one, while the Agilent Bio mAb NP1.7 SS provides the highest retention. Regarding the selectivity achieved between the main mAbs isoforms and their variants, the Thermo MabPac SCX column generally gave the highest selectivity. Finally, it was hard to rank columns in term of kinetic performance since their performance is strongly solute (mAb) and elution mode (pH or salt gradient) dependent. However, the highest resolution--in most cases--was observed on the strong cation exchanger YMC Bio Pro SP-F material.
阳离子交换色谱法（CEX）是一种成熟的单克隆抗体（mAb）表征策略。流动相条件的优化在文献中有详细描述，但缺乏关于用于治疗性蛋白质分析的CEX固定相的信息。本研究的目的是基于在pH梯度和盐梯度模式下使用各种治疗性单克隆抗体及其变体所实现的保留、选择性和分离能力，比较五种最先进的CEX固定相。本研究中考虑了Sepax Antibodix WCX-NP3、赛默飞世尔MAbPac SCX-10 RS、YMC BioPro SP-F、沃特世Protein-Pak Hi Res SP和安捷伦Bio mAb NP1.7 SS。在保留方面，YMC Bio Pro SP-F材料的保留能力最弱，而安捷伦Bio mAb NP1.7 SS的保留能力最强。关于主要单克隆抗体异构体及其变体之间实现的选择性，赛默飞世尔MabPac SCX柱通常具有最高的选择性。最后，由于柱的性能强烈依赖于溶质（单克隆抗体）和洗脱模式（pH或盐梯度），因此很难根据动力学性能对柱进行排名。然而，在大多数情况下，在强阳离子交换剂YMC Bio Pro SP-F材料上观察到了最高的分辨率。
