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解读单克隆抗体轻链染色学峰值分裂的LC-MS方法

Yanjun Liu¹, Zac VanAernum², Yue Zhang^3,4

¹ProtaGene US, Inc. was Formerly BioAnalytix Inc., 4 Burlington Woods Dr., Burlington, MA, 01803, USA. yanjun.liu@protagene.com.

Pharmaceutical research

|November 7, 2023

概括

由于酸异体化而导致的单克隆抗体 (mAb) 降解被使用LC-MS确定. 在CDR中的这种isoAsp形成降低了治疗功效,强调了需要强有力的质量控制.

科学领域:

生物制药学分析
蛋白质化学蛋白质化学
分析化学分析化学

背景情况:

单克隆抗体 (mAbs) 容易降解,影响它们的效力和疗效.
从本地mAbs中区分退化形式是具有挑战性的,但对于质量控制至关重要.
了解产品质量属性 (PQA) 对于优化制造和配方至关重要.

研究的目的:

开发一种可通用的液体染色学-质谱学 (LC-MS) 方法,用于在mAbs.中量化异酸盐 (isoAsp).
为了确定在mAb分析中观察到的峰值分裂的根本原因.
为了将isoAsp形成与mAb强度的变化相关联.

主要方法:

利用逆相色谱,观察热应激mAbs.中的峰值分裂.
采用多种LC-MS方法,包括补充HPLC列和各种MS/MS分离技术.
进行了酶性消化并收集了分裂峰值以确认ISOAsp丰富.
使用酶相关免疫吸收试验 (ELISA) 测量了mAb功效.

主要成果:

在mAbs的峰值分裂被归因于光链的互补性决定区域 (CDR) 中的酸异构化.
LC-MS分析证实了分裂峰的相同质量,在早期排泄峰上有isoAsp的丰富.
热应激样本显示强度下降与CDR中ISOAsp水平增加相关.

结论:

成功开发了一种可通用的LC-MS方法,用于在mAbs.中识别和量化含有isoAsp的.
这种方法有助于理解mAb降解及其对功效的影响.
这些发现支持使用LC-MS进行生物制品的全面PQA评估.

关键词:

在 LCMS 中使用.确定互补性的地区 (CDR)关键质量属性 (CQA) 是一个关键质量属性.异构化的异构化一个单克隆抗体 (mab)

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