Comprehensive analysis of N-glycans in IgG purified from ferrets with or without influenza A virus infection

Abstract

This work is supported by Food and Drug Administration intramural funds. This project was supported in part by an appointment to the Internship/ Research Participation Program at the Office of Biotechnology Products, Food and Drug Administration, administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the Department of Energy and Food and Drug Administration. The authors declare that they have no conflicts of interest with the contents of this article. The views expressed in this article are those of the authors, and do not necessarily reflect the official policy or position of the U.S. Food and Drug Administration and the Dept. of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. This article contains Figs. S1-S14 and Tables S1-S3.

Biophysical Evaluation of Rhesus Macaque Fc Gamma Receptors Reveals Similar IgG Fc Glycoform Preferences to Human Receptors

“…In contrast, the presence or absence of bisecting GlcNAc did not have a statistically significant effect on the binding affinity of any RM FcgR tested (Figures 2B, 3). While a prior study suggested that the presence of bisecting GlcNAc resulted in improved effector function in the context of human FcgR (52), other work has suggested that these observations were instead driven by variable fucosylation (51,53). The observation that bisected glycans cannot subsequently become fucosylated appears to have resulted in some confounding of cause and effect with respect to the role of bisection (54).…”

“…Because the effect of variable fucose content is of clinical relevance in both natural immune responses (19)(20)(21)(22)(23)(24) and in optimization of antibody therapy (31,53), we further probed this aspect of RM receptor binding profiles with additional antibody specificities. Rhesusized antibody to CD20 engineered to lack fucose (Supplemental Figure 2) showed improved binding affinity to RM FcgRIII, as did a Dual Variable Domain (DVD) format bispecific (Figure 5) that was similarly modified to reduce fucose content (Supplemental Figure 3).…”

N-Glycosylation of mollusk hemocyanins contributes to their structural stability and immunomodulatory properties in mammals

“…Indeed, glycosylations reduce chemical and physical protein instabilities as they increase the internal noncovalent forces and the solvent-accessible surface area, among other parameters. The effects of glycosylation on protein stability are highly sensitive to the location of the glycosylation site in the tridimensional structure of the proteins (47–49). In the case of hemocyanins, some N- glycans are located at the C-terminal end of FUs, at the interface between subunits (16).…”

Glycomics and glycoproteomics: Approaches to address isomeric separation of glycans and glycopeptides

“…The native glycan is made less hydrophilic by the substitution of the hydrogen atoms in all hydroxyl, N ‐acetyl, and carboxylic acid moieties for methyl groups. Glycan permethylation is particularly efficient for the separation of isomers and, therefore, it is the most used derivatization technique for glycome analysis [3,7,11,16,20,82,84,85,95,96,138–142]. Otherwise, the use of permethylation in glycopeptides is not an easy task; to date, there is no reliable proof of its application [143].…”