Fc glycan sialylation of biotherapeutic monoclonal antibodies has limited impact on antibody‐dependent cellular cytotoxicity

Abstract

It has been well-documented that the terminal sugars of Fc glycans can play a critical role in the safety and efficacy of therapeutic monoclonal antibodies (mAbs). However, many of the effects of highly heterogeneous Fc glycan structures have yet to be fully characterized. Different glycosylation patterns can affect Fcdependent activities such as the ability of mAbs to bind Fcγ receptors (FcγRs) on the effector cell surface, which is critical to immune effector functions such as antibody dependent cellular cytotoxicity (ADCC).Previous studies on the impact of sialic acid in the Fc glycan on ADCC has not resulted in consistent conclusions. In our study we tested sialic acid enriched species from a chimeric murine/human kappa light chain IgG1 (mAb1) with known FcγRIIIa binding and ADCC activities. These enriched species contained up to a four-fold increase in sialic acid-containing glycans relative to the typical levels present in therapeutic mAbs, along with other attributes such as oxidized and deamidated species. The ADCC analysis of sialylated

Cell-based production of Fc-GlcNAc and Fc-alpha-2,6 sialyl glycan enriched antibody with improved effector functions through glycosylation pathway engineering

“…1). Furthermore, the terminal sialylation with 2-6 linkage, not the 2-3 linkage, was introduced as it was found to enhance receptor binding 31 and improve ADCC and vaccinal effect [23][24]32 . Although the mRNA of human 2-3 sialyltransferases could be detected in 293T and CHO cells 18,33 , we did not observe any obvious sialylation on the antibody glycan from these cell lines [18][19]29 (Fig.…”

International standards for monoclonal antibodies for assessing the biological activity of medicines: A status update

“…К первой относятся методы оценки связывания, такие как твердофазный иммуноферментный анализ (ИФА, ELISA), метод проточный цитофлуориметрии, методы флуоресцентного резонансного переноса энергии и поверхностного плазмонного резонанса, позволяющие охарактеризовать функции, опосредованные структурой МкАТ [11][12][13][14]. С помощью перечисленных методов проводится изучение связывания МкАТ с мишенью или с рецепторами клеточной поверхности, такими как FcγR и FcRn, а также с компонентами системы комплемента для оценки Fc-опосредованной цитотоксичности [15][16][17]. Методы in vitro, наиболее часто применяемые при изучении биологических свойств препаратов МкАТ, представлены в таблице 1.…”

“…В отчетах о результатах международных исследований по разработке и изучению МСО этанерцепта, ритуксимаба, инфликсимаба, ада лимумаба, бевацизумаба, трастузумаба 16 и цетуксимаба 17 отмечается, что применение в соответствующих биологических тестах препаратовкандидатов в МСО, в отличие от внутренних СО производителей, способствовало гармонизации оценки биологической активности между лабораториями даже при использовании нескольких методик, основанных на разных принципах [25,28,[32][33][34].…”

Structure-Specific N-Glycoproteomics Characterization of NIST Monoclonal Antibody Reference Material 8671

“…Microheterogeneity develops its importance in mAbs. For example, mAbs with terminal sialic acids have a longer half-life in the case of an asialoglycoprotein receptor and may influence the function of ADCC and CDC; − on the other hand, mAbs with terminal mannose have a shorter half-life and weaker ADCC . The biggest influence comes from core fucose in ADCC, which inhibits the effector, whereas the bisect GlcNAc has an opposite effect .…”