The impact of N-glycosylation on conformation and stability of immunoglobulin Y from egg yolk

- IgY molecule became more flexible and disordered after removal of N-glycan.
- Trp residues of IgY were buried in more hydrophobic environment after enzymolysis.
- Storage stability of IgY reduced after deglycosylation based on SEC-HPLC analysis.
- Deglycosylated IgY exhibited less resistance to GdnHCl-induced unfolding.
- After deglycosylation, IgY was more sensitive to pepsin.

Immunoglobulin Y (IgY) is a new therapeutic antibody, and its applications in industry are very broad. To provide insight into the effects of N-glycosylation on IgY, its conformation and stability were studied. In this research, IgY was extracted from egg yolk and then digested by peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase. SDS-PAGE and infrared absorption spectrum showed that carbohydrates were distinctly reduced after enzymolysis. The circular dichroism spectrum indicated that the IgY molecule became more flexible and disordered after removal of N-glycan. The fluorescence intensity revealed that Trp residues were buried in a more hydrophobic environment after disposal of N-glycan. Storage stability decreased with the removal of oligosaccharide chains based on size-exclusion chromatography analysis. Deglycosylated IgY exhibited less resistance to guanidine hydrochloride-induced unfolding. After deglycosylation, IgY was more sensitive to pepsin. Therefore, N-glycosylation played an important role in the maintenance of the structure and stability of IgY.