Naslov
Lectin and Liquid Chromatography-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Autori
Petrović, Tea ; Trbojević-Akmačić, Irena

Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, pregledni

Knjiga
Antibody Glycosylation

Urednik/ci
Pezer, Marija

Izdavač
Springer

Grad
Cham

Godina
2021

Raspon stranica
29-72

ISSN
1664-431X

Ključne riječi
Critical quality attribute ; Glycosylation analysis ; Immunoglobulin ; Lectin chromatography ; Lectin microarrays ; Lectins ; Liquid chromatography ; Mass spectrometry ; N-glycans ; O-glycans

Sažetak
Immunoglobulin (Ig) glycosylation has been shown to dramatically affect its structure and effector functions. Ig glycosylation changes have been associated with different diseases and show a promising biomarker potential for diagnosis and prognosis of disease advancement. On the other hand, therapeutic biomolecules based on structural and functional features of Igs demand stringent quality control during the production process to ensure their safety and efficacy. Liquid chromatography (LC) and lectin-based methods are routinely used in Ig glycosylation analysis complementary to other analytical methods, e.g., mass spectrometry and capillary electrophoresis. This chapter covers analytical approaches based on LC and lectins used in low- and high-throughput N- and O-glycosylation analysis of Igs, with the focus on immunoglobulin G (IgG) applications. General principles and practical examples of the most often used LC methods for Ig purification are described, together with typical workflows for N- and O-glycan analysis on the level of free glycans, glycopeptides, subunits, or intact Igs. Lectin chromatography is a historical approach for the analysis of lectin-carbohydrate interactions and glycoprotein purification but is still being used as a valuable tool in Igs purification and glycan analysis. On the other hand, lectin microarrays have found their application in the rapid screening of glycan profiles on intact proteins.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Interdisciplinarne prirodne znanosti

POVEZANOST RADA