结合线上和线下二维液体染色学和质谱的组合,以全面描述mAb电荷变体,并为快速过程开发提供精确的指令
Xiaoqing Jin1, Bingfang He1,2
1College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
International journal of molecular sciences
|October 28, 2023
概括
这项研究引入了一种快速的2D-LC-MS方法,以精确识别单克隆抗体 (mAb) 电荷变异. 该平台通过为流程优化提供快速,可操作的见解来加速治疗开发.
科学领域:
- 生物制药学分析分析
- 蛋白质的表征 蛋白质的表征
- 分析化学是一种分析化学.
背景情况:
- 单克隆抗体 (mAbs) 是关键的治疗药物,但它们的充电变异构成质量属性挑战.
- 描述mAb电荷变体的传统方法是复杂的,耗时的,劳动密集的.
研究的目的:
- 开发一个全面和快速的平台来表征mAb充电变体.
- 使用先进的分析技术,为生物工艺开发提供精确的指令.
主要方法:
- 结合线上和线下二维液态染色学与质谱学 (2D-LC-MS) 的结合.
- 在完整的蛋白质和水平上进行分析,以确定特定的修改.
- 使用了少量样品 (0.6 mg mAb) 和短分析时间 (在6小时内).
主要成果:
- 成功识别了6个不同的充电变体峰值在mAb1.
- 离线的2D-LC-MS确定了除化,氧化和C端K损失.
- 在线2D-LC-MS检测到谷甲和糖化,揭示了由于预处理器件而在线分析中错过的修改.
结论:
- 开发的在线和离线2D-LC-MS平台提供了全面,精确和快速的mAb充电变体的表征.
- 这种方法可以实现精确的工艺开发,专注于特定的修改,如针对性细胞培养优化的谷氨酸化.
- 该平台通过在数小时内提供可操作数据来加速治疗开发.
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