IgE Reactivity of the Dog Lipocalin Allergen Can f 4 and the Development of a Sandwich ELISA for Its Quantification

Allergy, Asthma & Immunology Research
M Rytkönen-NissinenT Virtanen

Abstract

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. 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). 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 e...Continue Reading

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Citations

Feb 15, 2016·Current Allergy and Asthma Reports·Julia ZimmerSusanne Kaul
Apr 26, 2016·Canine Genetics and Epidemiology·Christina BreitenbuecherAnita M Oberbauer
May 18, 2016·Current Allergy and Asthma Reports·Anna PomésSabina Wünschmann
Mar 20, 2019·Allergy·Anna WintersandHans Grönlund
Jan 15, 2021·The Journal of Allergy and Clinical Immunology·Ann-Marie M SchoosMagnus P Borres
Dec 6, 2021·The Journal of Allergy and Clinical Immunology·Céline VandammeTuomas Virtanen

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Datasets Mentioned

BETA
KF192077

Methods Mentioned

BETA
ELISA
Protein Assay
electrophoresis
reversed phase chromatography
dot blot
ELISAs
size exclusion chromatography

Software Mentioned

ImageJ Microarray
GraphPad Prism
SPSS SPSS
ImageJ
Mascot

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