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A Human IgE Antibody Binding Site on Der p 2 for the Design of a Recombinant Allergen for Immunotherapy.
- Source :
-
Journal of immunology (Baltimore, Md. : 1950) [J Immunol] 2019 Nov 01; Vol. 203 (9), pp. 2545-2556. Date of Electronic Publication: 2019 Sep 25. - Publication Year :
- 2019
-
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 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.<br /> (Copyright © 2019 by The American Association of Immunologists, Inc.)
- Subjects :
- Antibodies, Monoclonal chemistry
Antigens, Dermatophagoides chemistry
Arthropod Proteins chemistry
Crystallography, X-Ray
Epitopes immunology
Humans
Magnetic Resonance Spectroscopy
Mutagenesis, Site-Directed
Protein Conformation
Recombinant Proteins immunology
Antibodies, Monoclonal immunology
Antigens, Dermatophagoides immunology
Arthropod Proteins immunology
Binding Sites, Antibody
Desensitization, Immunologic methods
Immunoglobulin E immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1550-6606
- Volume :
- 203
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Journal of immunology (Baltimore, Md. : 1950)
- Publication Type :
- Academic Journal
- Accession number :
- 31554696
- Full Text :
- https://doi.org/10.4049/jimmunol.1900580