Immunoglobulin G glycoprofiles are unaffected by common bottom-up sample processing

Amez Martin, M.; Wuhrer, M.; Falck, D. (2020)

Article / Letter to editor

Immunoglobulin G (IgG) glycosylation is a key post-translational modification in regulating IgG function. It is therefore a prominent target for biomarker discovery and a critical quality attribute of antibody-based biopharmaceuticals. A common approach for IgG glycosylation analysis is the measurement of tryptic glycopeptides. Glycosylation stability during sample processing is a key prerequisite for an accurate and robust analysis yet has hitherto hardly been studied. Especially, acid hydrolysis of sialic acids may be a source for instability. Therefore, we investigated acid denaturation, centrifugal vacuum concentration, and glycopeptide storage regarding changes in the IgG glycosylation profile. Intravenous IgG was analyzed employing imaginable deviations from a reference method and stress conditions. All glycosylation features-sialylation, galactosylation, bisection, and fucosylation-remained unchanged for most conditions. Only with prolonged exposure to acidic conditions at 37 degrees C, sialylation decreased significantly and subtle changes occurred for galactosylation. Consequently, provided that long or intense heating in acidic solutions is avoided, sample preparation for bottom-up glycoproteomics does not introduce conceivable biases.

• glycoproteomics
• glycosylation stability
• immunoglobulin glycosylation
• acid hydrolysis
• glycopeptide storage

• All authors: Amez Martin, M.; Wuhrer, M.; Falck, D.
• Date: 2020-10-02
• Journal: Journal of Proteome Research
• Volume: 19
• Issue: 10
• Pages: 4158 - 4162
• DOI: doi:10.1021/acs.jproteome.0c00656
• Link: https://pubs.acs.org/doi/10.1021/acs.jproteome.0c00656