Comparative sedimentation equilibrium analysis of two IgG1 glycoforms: IgGCri and IgGWid.

Comparative sedimentation equilibrium analysis of two IgG1 glycoforms: IgGCri and IgGWid.
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DOI:
10.1007/s00249-023-01656-x
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发表时间:
2023-07
影响因子:
2
通讯作者:
Harding, Stephen E.
Harding, Stephen E.
中科院分区:
生物学4区
文献类型:
--
作者:
Hammad, Khalil Abu;Dinu, Vlad;MacCalman, Thomas E.;Pattem, Jacob;Goodall, Margaret;Gillis, Richard B.;Jefferis, Roy;Harding, Stephen E.

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主要使用沉降平衡分析和两个补充分析程序(SEDFIT-MSTAR 和 MULTISIG)来比较 IgG1 的两种不同糖型(IgG1Cri 和 IgG1Wid)的溶液特性。 IgGCri 在其 Fc 结构域上具有双触角复合型聚糖,其核心完全岩藻糖基化和部分唾液酸化,而在 IgGWid 上,它们是非岩藻糖基化、部分半乳糖基化和非唾液酸化。 IgGWid 也被 Fab 糖基化。尽管存在这些差异,SEDFIT-MSTAR 分析显示 IgGCri 的重均摩尔质量 Mw 相似,分别为 ~ (150 ± 5) kDa 和 IgGWid 的 ~ (154 ± 5) kDa,并且两种糖型均显示出存在一小部分二聚体的证据,这经 MULTISIG 分析以及支持性沉降速度测量的沉降系数分布证实。不同浓度下两种糖型的沉降平衡行为和沉降系数分布的接近性(主峰沉降系数为 ~ 6.4S)表明不同的糖基化特征对溶液中的摩尔质量(分子量)和构象没有显着影响。
The solution properties of two different glycoforms of IgG1 (IgG1Cri and IgG1Wid) are compared using primarily sedimentation equilibrium analysis with two complementary analysis routines: SEDFIT-MSTAR and MULTISIG. IgGCri bears diantennary complex-type glycans on its Fc domain that are fully core fucosylated and partially sialylated, whilst on IgGWid, they are non-fucosylated, partially galactosylated and non-sialylated. IgGWid is also Fab glycosylated. Despite these differences, SEDFIT-MSTAR analysis shows similar weight average molar masses Mw of ~ (150 ± 5) kDa for IgGCri and ~ (154 ± 5) kDa for IgGWid and both glycoforms show evidence of the presence of a small fraction of dimer confirmed by MULTISIG analysis and also by sedimentation coefficient distributions from supportive sedimentation velocity measurements. The closeness of the sedimentation equilibrium behaviour and sedimentation coefficient distributions with a main peak sedimentation coefficient of ~ 6.4S for both glycoforms at different concentrations suggest that the different glycosylation profiles do not significantly impact on molar mass (molecular weight) nor conformation in solution.
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