用CZE-MS对单克隆抗体进行充电变异分析,使用基于二甲基胺氨基乙烯-德克斯兰的连续多重离子聚合物层涂层
Alisa Höchsmann1,2, Laura Dhellemmes3, Laurent Leclercq3
1Faculty of Chemistry, Aalen University, Aalen, Germany.
Electrophoresis
|September 17, 2024
概括
这项研究引入了一种新的毛细管区电泳 (CZE) 方法,加上电喷射电离质谱法 (ESI-MS),用于分析单克隆抗体 (mAb) 电荷变异. 该技术可以精确地表征杂质,提高制药抗体的安全性和功能.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 制药科学 制药科学
背景情况:
- 药品单克隆抗体 (mAbs) 中杂质的特征对于确保药物的有效性和患者安全至关重要.
- 毛细管区电泳 (CZE) 对于分离mAb电荷变体是有效的,但其与质谱学的兼容性受到背景电解质的限制.
- 电子喷射电离质谱 (ESI-MS) 对于详细的杂质分析至关重要,但传统的CZE缓冲器阻碍了其应用.
研究的目的:
- 开发和验证CZE-ESI-MS方法,用于直接分离和表征完整的mAb电荷变体.
- 为了克服CZE背景电解质和ESI-MS.之间的不兼容问题.
- 为了证明开发的方法用于分析各种mAb蛋白形的广泛适用性.
主要方法:
- 开发了一种新的阴离子毛细血管涂层,使用连续多个离子聚合物层的二甲基氨基乙基-德克斯-多聚 ( styrene sulfonate).
- 酸性背景电解质 (BGE) 与涂层毛细血管一起使用,以实现最佳分离.
- 该方法使用USP mAb003进行了验证,并与其他治疗抗体如infliximab,adalimumab和trastuzumab进行了进一步测试.
主要成果:
- 开发的CZE-ESI-MS方法成功地分离了USP mAb003.3.的C端 lysine变体,酸性变体和单糖化形式.
- 阴离子涂层产生了反向电流 (EOF),允许分离具有微妙移动性差异的变体.
- 该方法在多种治疗性单克隆抗体中显示出普遍适用性,包括NISTmAB,阿达利穆马布和特拉斯图祖马布.
结论:
- 新的CZE-ESI-MS技术为mAb电荷变体和蛋白形体的全面表征提供了一个强大的平台.
- 这种方法提高了单克隆抗体开发中的质量控制和杂质分析能力.
- 证明的多功能性表明它有可能用于对各种治疗抗体的常规分析.
关键词:
毛细管涂层是毛细管的涂层毛细血管电泳 毛细血管电泳阴离子聚电解质多层多层.完好无损的抗体分离.质谱测量质谱测量质谱测量质谱测量质量测量质谱测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量更多相关视频
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