Immunoglobulin G specifically binding plant N-glycans with high affinity could be generated in rabbits but not in mice.

Abstract:

:Xylosylated and core alpha1,3-fucosylated N-glycans from plants are immunogenic, and they play a still obscure role in allergy and in the field of plant-made protein pharmaceuticals. We immunized mice to generate monoclonal antibodies (mAbs) binding plant N-glycans specifically via the epitope containing either the xylose or the core alpha1,3-fucose residue. Splenocytes expressing N-glycan-specific antibodies derived from C57BL/6 mice previously immunized with plant glycoproteins were preselected by cell sorting to generate hybridoma lines producing specific antibodies. However, we obtained only mAbs unable to distinguish fucosylated from xylosylated N-glycans and reactive even with the pentasaccharide core Man3GlcNAc2. In contrast, immunization of rabbits yielded polyclonal sera selectively reactive with either fucosylated or xylosylated N-glycans. Purification of these sera using glyco-modified neoglycoproteins coupled to a chromatography matrix provided polyclonal sera suitable for affinity determination. Surface plasmon resonance measurements using sensor chips with immobilized glyco-modified transferrins revealed dissociation constants of around 10(-9) M. This unexpectedly high affinity of IgG antibodies toward carbohydrate epitopes has repercussions on our conception of the binding strength and significance of antiglycan IgE antibodies in allergy.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Jin C,Bencúrová M,Borth N,Ferko B,Jensen-Jarolim E,Altmann F,Hantusch B

doi

10.1093/glycob/cwj071

subject

Has Abstract

pub_date

2006-04-01 00:00:00

pages

349-57

issue

4

eissn

0959-6658

issn

1460-2423

pii

cwj071

journal_volume

16

pub_type

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