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在天然丰度下使用NMR对糖基化Fc的结构性扰动进行特定地点的表征

Béatrice Vibert^1,2, Sarah Nguyen^3,4,5, Faustine Henot^6,7,3

¹Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale (IBS), 38044, Grenoble, France. beatrice.vibert@ibs.fr.

Journal of biomolecular NMR

|February 17, 2026

概括

研究人员在治疗抗体的Fc片段中分配了甲基共振,从而实现了详细的结构分析. 这一突破提高了单克隆抗体 (mAbs) 的质量控制,允许精确检测结构变化.

科学领域:

生物化学生物化学
结构生物学结构生物学
分析化学分析化学

背景情况:

单克隆抗体 (mAbs) 是关键的治疗药物,需要严格的质量控制,以确保结构完整性和批量一致性.
甲基2D核磁共振对于探测自然同位素丰度下的mAb结构是有价值的,有助于批量对批量进行比较.
对抗体的结晶片段 (Fc) 缺乏全面的甲基共振赋值,阻碍了详细的结构分析.

研究的目的:

为了实现免疫球蛋白G1 (IgG1) 抗体的Fc片段的甲基组分配.
为了使原子层面的结构洞察到使用NMR治疗抗体.
推进mAb生产的质量控制方法.

主要方法:

在NMR实验中利用细胞自由表达和战略同位素标记.
在非糖化Fc片段上进行高质量的2D和3DNMR.
转移了从中国仓鼠卵巢 (CHO) 生产的糖化Fc片段的自然丰富的mAb的分配.

主要成果:

在非糖化Fc片段中成功分配了94%的甲基共振.
从CHO产生的mAb.Ab中获得一个糖基化Fc片段的83%的分配覆盖率.
在原子分辨率下研究甲氨酸氧化对Fc结构的影响.

结论:

Fc片段的甲基组分配将二维甲基NMR指纹转化为一个强大的质量控制工具.
能够直接对自然丰富的mAbs进行光谱比较,以便在没有同位素标记的情况下检测和定位修改.
在开发和制造过程中提供了一种可靠的方法来监测治疗性抗体的结构完整性.

关键词:

在Fc Fc Fc 在 IgG1 mAb 的情况下,甲基组是甲基组的组成部分.核磁共振 (NMR) 的分配方式质量控制质量控制

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