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A Human IgE Antibody Binding Site on Der p 2 for the Design of a Recombinant Allergen for Immunotherapy

Glesner, Jill ; Kapingidza, A. Brenda ; Godzwon, Magdalena LU ; Offermann, Lesa R. ; Mueller, Geoffrey A. ; DeRose, Eugene F. ; Wright, Paul ; Richardson, Crystal M. ; Woodfolk, Judith A. and Vailes, Lisa D. , et al. (2019) In Journal of immunology 203(9). p.2545-2556
Abstract

Der p 2 is one of the most important allergens from the house dust mite Dermatophagoides pteronyssinus Identification of human IgE Ab binding epitopes can be used for rational design of allergens with reduced IgE reactivity for therapy. Antigenic analysis of Der p 2 was performed by site-directed mutagenesis based on the x-ray crystal structure of the allergen in complex with a Fab from the murine IgG mAb 7A1 that binds an epitope overlapping with human IgE binding sites. Conformational changes upon Ab binding were confirmed by nuclear magnetic resonance using a 7A1-single-chain variable fragment. In addition, a human IgE Ab construct that interferes with mAb 7A1 binding was isolated from a combinatorial phage-display library... (More)

Der p 2 is one of the most important allergens from the house dust mite Dermatophagoides pteronyssinus Identification of human IgE Ab binding epitopes can be used for rational design of allergens with reduced IgE reactivity for therapy. Antigenic analysis of Der p 2 was performed by site-directed mutagenesis based on the x-ray crystal structure of the allergen in complex with a Fab from the murine IgG mAb 7A1 that binds an epitope overlapping with human IgE binding sites. Conformational changes upon Ab binding were confirmed by nuclear magnetic resonance using a 7A1-single-chain variable fragment. In addition, a human IgE Ab construct that interferes with mAb 7A1 binding was isolated from a combinatorial phage-display library constructed from a mite-allergic patient and expressed as two recombinant forms (single-chain Fab in Pichia pastoris and Fab in Escherichia coli). These two IgE Ab constructs and the mAb 7A1 failed to recognize two Der p 2 epitope double mutants designed to abolish the allergen-Ab interaction while preserving the fold necessary to bind Abs at other sites of the allergen surface. A 10-100-fold reduction in binding of IgE from allergic subjects to the mutants additionally showed that the residues mutated were involved in IgE Ab binding. In summary, mutagenesis of a Der p 2 epitope defined by x-ray crystallography revealed an IgE Ab binding site that will be considered for the design of hypoallergens for immunotherapy.

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type
Contribution to journal
publication status
published
subject
in
Journal of immunology
volume
203
issue
9
pages
12 pages
publisher
American Association of Immunologists
external identifiers
  • scopus:85073666428
  • pmid:31554696
ISSN
1550-6606
DOI
10.4049/jimmunol.1900580
language
English
LU publication?
yes
id
e059e614-144a-4792-9ba9-41c4662caa0a
date added to LUP
2019-10-29 14:51:16
date last changed
2024-06-12 03:32:31
@article{e059e614-144a-4792-9ba9-41c4662caa0a,
  abstract     = {{<p>Der p 2 is one of the most important allergens from the house dust mite Dermatophagoides pteronyssinus Identification of human IgE Ab binding epitopes can be used for rational design of allergens with reduced IgE reactivity for therapy. Antigenic analysis of Der p 2 was performed by site-directed mutagenesis based on the x-ray crystal structure of the allergen in complex with a Fab from the murine IgG mAb 7A1 that binds an epitope overlapping with human IgE binding sites. Conformational changes upon Ab binding were confirmed by nuclear magnetic resonance using a 7A1-single-chain variable fragment. In addition, a human IgE Ab construct that interferes with mAb 7A1 binding was isolated from a combinatorial phage-display library constructed from a mite-allergic patient and expressed as two recombinant forms (single-chain Fab in Pichia pastoris and Fab in Escherichia coli). These two IgE Ab constructs and the mAb 7A1 failed to recognize two Der p 2 epitope double mutants designed to abolish the allergen-Ab interaction while preserving the fold necessary to bind Abs at other sites of the allergen surface. A 10-100-fold reduction in binding of IgE from allergic subjects to the mutants additionally showed that the residues mutated were involved in IgE Ab binding. In summary, mutagenesis of a Der p 2 epitope defined by x-ray crystallography revealed an IgE Ab binding site that will be considered for the design of hypoallergens for immunotherapy.</p>}},
  author       = {{Glesner, Jill and Kapingidza, A. Brenda and Godzwon, Magdalena and Offermann, Lesa R. and Mueller, Geoffrey A. and DeRose, Eugene F. and Wright, Paul and Richardson, Crystal M. and Woodfolk, Judith A. and Vailes, Lisa D. and Wünschmann, Sabina and London, Robert E. and Chapman, Martin D. and Ohlin, Mats and Chruszcz, Maksymilian and Pomés, Anna}},
  issn         = {{1550-6606}},
  language     = {{eng}},
  number       = {{9}},
  pages        = {{2545--2556}},
  publisher    = {{American Association of Immunologists}},
  series       = {{Journal of immunology}},
  title        = {{A Human IgE Antibody Binding Site on Der p 2 for the Design of a Recombinant Allergen for Immunotherapy}},
  url          = {{http://dx.doi.org/10.4049/jimmunol.1900580}},
  doi          = {{10.4049/jimmunol.1900580}},
  volume       = {{203}},
  year         = {{2019}},
}