IgE Reactivity of the Dog Lipocalin Allergen Can f 4 and the Development of a Sandwich ELISA for Its Quantification.
10.4168/aair.2015.7.4.384
- Author:
Marja RYTKONEN-NISSINEN
1
;
Soili SAARELAINEN
;
Jukka RANDELL
;
Jukka HAYRINEN
;
Nisse KALKKINEN
;
Tuomas VIRTANEN
Author Information
1. Department of Clinical Microbiology, Institute of Clinical Medicine and Biocenter Kuopio, University of Eastern Finland, Kuopio Campus, Finland. Marja.Rytkonen-Nissinen@uef.fi
- Publication Type:Original Article
- Keywords:
Allergen;
Can f 4;
Canis familiaris;
dog;
IgE;
lipocalin
- MeSH:
Allergens;
Animals;
Antibodies, Monoclonal;
Chromatography, Affinity;
Dander;
Dogs*;
Enzyme-Linked Immunosorbent Assay*;
Humans;
Hypersensitivity;
Immunoblotting;
Immunoglobulin E*;
Lipocalins*;
Skin
- From:Allergy, Asthma & Immunology Research
2015;7(4):384-392
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Divergent results on the IgE reactivity of dog-allergic subjects to Can f 4 have been reported. The aim of this study was to evaluate the significance of Can f 4 in dog allergy and to develop an immunochemical method for measuring Can f 4 content in environmental samples. METHODS: We purified the natural dog allergen Can f 4 from a dog dander extract by monoclonal antibody-based affinity chromatography and generated its variant in a recombinant form. Sixty-three dog-allergic patients and 12 nonallergic control subjects were recruited in the study. The IgE-binding capacity of natural Can f 4 and its recombinant variant was assessed by ELISA, immunoblotting, and skin prick tests (SPT). RESULTS: Eighty-one percent of the dog-allergic patients showed a positive result to the immunoaffinity-purified natural Can f 4 in IgE ELISA, but only 46% in IgE immunoblotting. Respective results with the recombinant Can f 4 variant were 54% and 49%. SPT results reflected those obtained in ELISA and immunoblotting. The overall IgE reactivity of the immunoaffinity-purified natural Can f 4 was found to depend strongly on the integrity of the allergen's conformation. A sandwich ELISA based on monoclonal antibodies was found to be functional for measuring Can f 4 in environmental samples. CONCLUSIONS: Can f 4 is a major allergen of dog together with Can f 1 and Can f 5. In combination with other dog allergens, it improves the reliability of allergy tests in dog allergy.
