IgE Reactivity of the Dog Lipocalin Allergen Can f 4 and the Development of a Sandwich ELISA for Its Quantification |
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Authors: | Marja Rytk?nen-Nissinen Soili Saarelainen Jukka Randell Jukka H?yrinen Nisse Kalkkinen Tuomas Virtanen |
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Institution: | 1.Department of Clinical Microbiology, Institute of Clinical Medicine and Biocenter Kuopio, University of Eastern Finland, Kuopio Campus, Finland.;2.Institute of Dentistry, School of Medicine, University of Eastern Finland, Kuopio Campus, Finland.;3.Department of Pulmonary Diseases, Kuopio University Hospital, Kuopio, Finland.;4.Institute of Biomedicine, University of Eastern Finland, Kuopio Campus, Finland.;5.Institute of Biotechnology, Viikki Biocenter, University of Helsinki, Finland. |
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Abstract: | PurposeDivergent 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.MethodsWe 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).ResultsEighty-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.ConclusionsCan 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. |
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Keywords: | Allergen Can f 4 Canis familiaris dog IgE lipocalin |
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