He Xiaoping Z, Que Amy H, Mo Jim J
Global Biologics, Pfizer Global Research & Development, Pfizer Inc., Chesterfield, MO 63017, USA.
Electrophoresis. 2009 Mar;30(5):714-22. doi: 10.1002/elps.200800636.
An analytical method using the imaged CE (iCE) technology has been developed and validated for measuring contents of charge variants for mAb molecules. The method could generate similar information that require both conventional IEF electrophoresis and ion exchange liquid chromatography to generate as an iCE analysis would produce both the pI of the molecules and quantitative contents of charge variants. Thus, it offers unique advantages over the IEF and ion exchange methods in terms of identification, separation and quantitation of charge variants. The data presented in this study demonstrated that the iCE method is suitable not only for research purposes but also for quality control purpose of mAb clinical or commercial manufacture as the technique can be validated and possesses sufficient robustness. The developed generic iCE method has been tested for a wide range of therapeutic mAbs and proved its suitability for multiple mAb molecules. The tested mAbs are a group of molecules with a wide range of charge compositions (acidic species ranging from 10 to 70%) and a diverse range of pI of 6.9-9.6. Developing platform analytical technologies is an efficient way to meet the demand of rapid growth of therapeutic mAb candidates under clinical and preclinical development. The iCE technology is a good candidate to become a platform method as it could cover a broad range of pH gradient. The experiences learned during the developmental process would provide important and valuable information to the biotech industry for the evaluation of charge heterogeneity of mAbs in terms of release testing, characterization, stability study, process development support and comparability study.
一种使用成像毛细管电泳（iCE）技术的分析方法已被开发并验证，用于测定单克隆抗体（mAb）分子电荷变体的含量。该方法能够得出与传统等电聚焦（IEF）电泳和离子交换液相色谱法相同的信息，因为iCE分析既可以得出分子的等电点（pI），也可以得出电荷变体的定量含量。因此，在电荷变体的鉴定、分离和定量方面，它比IEF和离子交换方法具有独特优势。本研究中呈现的数据表明，iCE方法不仅适用于研究目的，也适用于mAb临床或商业生产的质量控制，因为该技术可以进行验证且具有足够的稳健性。已开发的通用iCE方法已针对多种治疗性mAb进行了测试，并证明其适用于多种mAb分子。所测试的mAb是一组电荷组成范围广泛（酸性物种范围为10%至70%）且等电点在6.9 - 9.6之间变化多样的分子。开发平台分析技术是满足临床和临床前开发中治疗性mAb候选药物快速增长需求的有效途径。iCE技术有望成为一种平台方法，因为它可以覆盖广泛的pH梯度范围。在开发过程中获得的经验将为生物技术行业在mAb电荷异质性的释放测试、表征、稳定性研究、工艺开发支持和可比性研究等方面的评估提供重要且有价值的信息。
