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通过共价标记和质谱法对双特异抗原结合生物治疗药物的构造变化进行映射

Arnik Shah^1,2, Dipanwita Batabyal³, Dayong Qiu¹

¹Amgen Inc., Cambridge, MA, 02141, USA.

Journal of pharmaceutical analysis

|September 12, 2024

概括

两种共价标签方法,二甲基碳酸盐 (DEPC) 标签和蛋白质的快速光氧化 (FPOP),有效地监测了双特异抗原结合生物疗法 (BABB) 的微妙结构变化. 这些技术提供了对生物治疗结构稳定性和产品质量的更深入的见解.

科学领域:

生物化学生物化学
蛋白质化学蛋白质化学
免疫治疗是一种免疫疗法.

背景情况:

生物治疗药物的高阶结构 (HOS) 对功能和相关性至关重要.
双特异抗原结合生物疗法 (BABB) 是一种新型的免疫瘤疗法.
传统的生物物理技术,如近紫外线循环二极化 (NUV-CD),在检测微妙的结构变化方面存在局限性.

研究的目的:

评估二甲基碳酸盐 (DEPC) 标记和蛋白质的快速光氧化 (FPOP) 与质谱学 (MS) 结合,用于研究BABBs中的结构修饰.
评估这些共价标签方法在检测氨基酸残留水平上微妙的结构变化的灵敏度.
探索这些方法在本地和热应力条件下监测构造变化的实用性.

主要方法:

甲基碳酸盐 (DEPC) 标签与质谱学 (MS) 结合.
蛋白质的快速光氧化 (FPOP) 与质谱学 (MS) 相结合.
在本地和热应力条件下对标签吸收的分析,通过绑定测试得到证实.

主要成果:

DEPC标记和FPOP-MS检测到BABBs的抗原结合域中的微妙结构变化.
这些变化在原生和热应力条件下都被观察到.
标记吸收变化与结合效应相关,并提供了对形状稳定性的见解.

结论:

与MS相结合的共价标记方法 (DEPC和FPOP) 是用于表征BABB结构的强大工具.
这些技术揭示了传统方法 (如NUV-CD) 无法检测到的形状变化.
开发的方法可以有效地监测生物治疗结构稳定性,影响产品质量并指导新疗法的开发.

关键词:

共价标签/足迹标签/足迹标签甲基甲基碳酸盐 (Diethylpyrocarbonate) 是一种碳酸盐.蛋白质的快速光氧化发生在蛋白质中.更高阶结构的特征化描述.液体染色学-质谱学液体染色学

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