通过液体染色学-质谱学进行治疗抗体碎片识别的系统工作流程
Xiaoxu Zhang1, Yanpeng Xu1, Liqi Xie1
1Analytical Science and Development Department, Henlius Biologics Co., Ltd., 5155 Guangfulin Road, Shanghai 201616, China.
Journal of the American Society for Mass Spectrometry
|November 11, 2024
概括
识别抗体分裂部位对于治疗开发至关重要. 这项研究提出了一种新的工作流程,使用多种技术来精确确定分裂部位,确保抗体的完整性和功能.
科学领域:
- 生物制药学分析分析
- 蛋白质化学 蛋白质化学
- 分析化学是一种分析化学.
背景情况:
- 抗体碎片化在生物制药开发中很常见,影响了疗效.
- 准确识别分离点对于流程优化至关重要.
- 像CE-SDS这样的现有方法缺乏精确的碎片识别.
研究的目的:
- 开发和验证一种系统的工作流程,用于识别抗体碎片化裂解点.
- 为了改善抗体碎片的表征.
- 确保治疗抗体的生物功能和稳定性.
主要方法:
- 集成的工作流,结合直接识别,片段丰富和片段确认.
- 使用微尺度映射 (PM),标记PM的N-终端和MW和PM的SDS-PAGE内凝提取.
- 使用液体染色学-质谱学 (LC-MS) 进行表征.
主要成果:
- 在两个治疗性抗体 (双特异性和四价抗体) 中成功识别了分裂部位.
- 在双特异抗体的链接器上局部关键裂纹,影响功能.
- 在四价抗体中确定了一个特定的LC/HC裂变部位 (G105-R106),而直接LC-MS则失败了.
结论:
- 开发的系统工作流允许精确识别抗体分裂部位.
- 这种方法对制药实验室具有广泛的适用性.
- 对碎片化机制的更好理解有助于优化抗体开发过程.
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