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

Abu Hammad, Khalil, Dinu, Vlad, MacCalman, Thomas E., Pattem, Jacob ORCID: https://orcid.org/0000-0003-0276-7996, Goodall, Margaret, Gillis, Richard B., Jeferis, Roy and Harding, Stephen E. 2023. Comparative sedimentation equilibrium analysis of two IgG1 glycoforms: IgGCri and IgGWid. European Biophysics Journal 52 (4-5) , pp. 439-443. 10.1007/s00249-023-01656-x

PDF - Published Version Available under License Creative Commons Attribution. Download (1MB)

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 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.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Dentistry
Publisher:	Springer
Date of First Compliant Deposit:	15 September 2023
Date of Acceptance:	14 March 2023
Last Modified:	17 Apr 2024 14:39
URI:	https://orca.cardiff.ac.uk/id/eprint/162529

Actions (repository staff only)

Edit Item