Glycobiology Advance Access published online on December 21, 2005
Glycobiology, doi:10.1093/glycob/cwj071
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1 Department of Chemistry, University of Natural Resources and Applied Life Sciences (BOKU), 1190 Vienna, Austria; These authors contributed equally to this work
* To whom correspondence should be addressed. Xylosylated and core
Received November 8, 2005
Revised December 13, 2005
Accepted December 16, 2005
Article
Immunoglobulin G specifically binding plant N-glycans with high affinity could be generated in rabbits but not in mice
Chunsheng Jin 1,
Monika Bencúrová 1,
Nicole Borth 2,
Boris Ferko 2,
Erika Jensen-Jarolim 3,
Friedrich Altmann 4 *,
and
Brigitte Hantusch 5
2 Department of Biotechnology, University of Natural Resources and Applied Life Sciences (BOKU), 1190 Vienna, Austria
3 Institute of Pathophysiology, Medical University Vienna, 1090 Vienna, Austria
4 Department of Chemistry, University of Natural Resources and Applied Life Sciences (BOKU), 1190 Vienna, Austria
5 Clinical Institute of Pathology, Medical University Vienna, 1090 Vienna, Austria
Friedrich Altmann, E-mail: friedrich.altmann{at}boku.ac.at
Abstract 
1,3-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 binding plant N-glycans specifically via the epitope containing either the xylose- or the core 1,3-fucose-residue. Splenocytes expressing N-glycan-specific antibodies derived from C57BL/6 mice previously immunized with plant glycoproteins were pre-selected by cell sorting to generate hybridoma lines producing specific antibodies. However, we obtained only monoclonal antibodies 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 neo-glycoproteins 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 towards carbohydrate epitopes has repercussions on our conception of the binding strength and significance of anti-glycan IgE antibodies in allergy.
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