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

IF 2.2 4区生物学 Q3 BIOPHYSICS

European Biophysics Journal Pub Date : 2023-05-17 DOI:10.1007/s00249-023-01656-x

Khalil Abu Hammad, Vlad Dinu, Thomas E. MacCalman, Jacob Pattem, Margaret Goodall, Richard B. Gillis, Roy Jefferis, Stephen E. Harding

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引用次数: 1

Abstract

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 M_w 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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IgG1两种糖型:IgGCri和IgGWid的沉降平衡比较分析

使用主要的沉降平衡分析和两个互补的分析程序:SEDFIT-MSTAR和MULTISIG，比较了IgG1的两种不同糖型(IgG1Cri和IgG1Wid)的溶液性质。IgGCri在其Fc结构域上具有完全核心聚焦和部分唾液化的双链复合物型聚糖，而在IgGWid上，它们是非聚焦，部分半乳糖化和非唾液化。IgGWid也被Fab糖基化。尽管存在这些差异，SEDFIT-MSTAR分析显示，IgGCri和IgGWid的重量平均摩尔质量相似，分别为~(150±5)kDa和~(154±5)kDa。MULTISIG分析和支持沉降速度测量的沉降系数分布证实，这两种糖型都存在一小部分二聚体。两种糖型在不同浓度下的沉降平衡行为和沉降系数分布非常接近，沉降系数主峰为~ 6.4S，这表明不同的糖基化分布对溶液中的摩尔质量(分子量)和构象没有显著影响。

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来源期刊

European Biophysics Journal 生物-生物物理

CiteScore

4.30

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The journal publishes papers in the field of biophysics, which is defined as the study of biological phenomena by using physical methods and concepts. Original papers, reviews and Biophysics letters are published. The primary goal of this journal is to advance the understanding of biological structure and function by application of the principles of physical science, and by presenting the work in a biophysical context. Papers employing a distinctively biophysical approach at all levels of biological organisation will be considered, as will both experimental and theoretical studies. The criteria for acceptance are scientific content, originality and relevance to biological systems of current interest and importance. Principal areas of interest include: - Structure and dynamics of biological macromolecules - Membrane biophysics and ion channels - Cell biophysics and organisation - Macromolecular assemblies - Biophysical methods and instrumentation - Advanced microscopics - System dynamics.