BACKGROUND: Plant profilins have been reported as minor allergens. They are a well-known pan-allergen family responsible for cross-reactivity between plant-derived foods and pollens. Watermelon profilin has been reported to be a major allergen in watermelon (Citrullus lanatus).The aim of this study was to characterize recombinant watermelon profilin, confirming its reactivity for diagnostic purposes and the development of immunotherapy. METHODS: Native profilin was purified from watermelon extract by affinity chromatography using poly-L-proline. Recombinant His-tagged profilin was produced in Pichia pastoris yeast using pPICZαA vector and purified by metal chelate affinity chromatography. ELISA and immunoblot were carried out with sera from 17 watermelon-allergic patients. Biological activity was tested by the basophil activation test. RESULTS: Native profilin and recombinant profilin were purified and identified by mass spectrometry. Both show similar IgE reactivity in vitro and are biologically active. CONCLUSIONS: Similarities were found in the IgE-binding patterns and biological activity of recombinant profilin and native profilin. Recombinant profilin may be a powerful tool for specific diagnosis.
BACKGROUND: Plant profilins have been reported as minor allergens. They are a well-known pan-allergen family responsible for cross-reactivity between plant-derived foods and pollens. Watermelon profilin has been reported to be a major allergen in watermelon (Citrullus lanatus).The aim of this study was to characterize recombinant watermelon profilin, confirming its reactivity for diagnostic purposes and the development of immunotherapy. METHODS: Native profilin was purified from watermelon extract by affinity chromatography using poly-L-proline. Recombinant His-tagged profilin was produced in Pichia pastorisyeast using pPICZαA vector and purified by metal chelate affinity chromatography. ELISA and immunoblot were carried out with sera from 17 watermelon-allergicpatients. Biological activity was tested by the basophil activation test. RESULTS: Native profilin and recombinant profilin were purified and identified by mass spectrometry. Both show similar IgE reactivity in vitro and are biologically active. CONCLUSIONS: Similarities were found in the IgE-binding patterns and biological activity of recombinant profilin and native profilin. Recombinant profilin may be a powerful tool for specific diagnosis.
Authors: Lesa R Offermann; Caleb R Schlachter; Makenzie L Perdue; Karolina A Majorek; John Z He; William T Booth; Jessica Garrett; Krzysztof Kowal; Maksymilian Chruszcz Journal: J Biol Chem Date: 2016-05-26 Impact factor: 5.157
Authors: A Brenda Kapingidza; Sarah E Pye; Noah Hyduke; Coleman Dolamore; Swanandi Pote; Caleb R Schlachter; Scott P Commins; Krzysztof Kowal; Maksymilian Chruszcz Journal: Mol Immunol Date: 2019-07-18 Impact factor: 4.407
Authors: José Alberto Torres; Manuel de Las Heras; Aroa Sanz Maroto; Fernando Vivanco; Joaquín Sastre; Carlos Pastor-Vargas Journal: J Biol Chem Date: 2014-07-03 Impact factor: 5.157
Authors: Barbara Cases; Maria Dolores Ibañez; Jose Ignacio Tudela; Silvia Sanchez-Garcia; Pablo Rodriguez Del Rio; Eva A Fernandez; Carmelo Escudero; Enrique Fernandez-Caldas Journal: World Allergy Organ J Date: 2014-05-08 Impact factor: 4.084