Deakin University
Browse

File(s) under permanent embargo

Recombinant expression and epitope mapping of grass pollen allergens

conference contribution
posted on 1996-01-01, 00:00 authored by Cenk SuphiogluCenk Suphioglu, P M Smith, E K Ong, R B Knox, M B Singh
We have studied the expression of recombinant forms of Group 1 allergens from rye-grass and Bermuda grass pollens. Recombinant Lol p 1 expressed in bacteria bound serum IgE from allergic patients. Based on analysis of fragments of the Lol p 1 cDNA clone, the major IgE-reactive epitope has been mapped to the C-terminus. However, although SDS-denatured natural Cyn d 1 (from Bermuda grass) bound IgE, the full or partial recombinant proteins expressed in bacteria did not bind IgE. We have since expressed Cyn d 1 in the yeast Pichia pastoris and restored IgE binding. cDNA clones encoding two isoforms of Lol p 5, Lol p 5A and Lol p 5B, have been expressed in bacteria and resulting polypeptides show IgE-binding. Random fragments of these clones have been generated and when expressed as partial recombinant proteins in bacteria, allowed us to identify the major IgE-binding epitopes. The allergenic epitopes were localised towards the C-terminal half of the molecule. Although both isoforms shared similar IgE-reactive epitopes, Lol p 5B did not recognise the Lol p 5A-specific monoclonal antibody A7. At sequence level, there appear to be several amino acid differences between the antigenic epitopes of these two isoallergens. These results aid in the design of diagnostics and in grass pollen immunotherapy.

History

Event

Molecular Blology of Allergens and the AtopiC Immune Response. Conference (2nd : 1995 : Quebec City, Quebec)

Volume

409

Series

Advances in Experimental Medicine and Biology

Pagination

147 - 155

Publisher

Springer

Location

Quebec City, Quebec

Place of publication

Berlin, Germany

Start date

1995-02-18

End date

1995-02-22

ISSN

0065-2598

Language

eng

Publication classification

E1.1 Full written paper - refereed

Editor/Contributor(s)

Nathan Back, Irun Cohen, David Kritchevsky, Abel Lajtha, Rodolfo Paolett

Title of proceedings

New Horizons in allergy Immunotherapy : Proceedings of the 2nd Internatioanl conference on the Molecular Biology of Allergens and the Atopic Immune Response

Usage metrics

    Research Publications

    Categories

    No categories selected

    Keywords

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC