SCOPE: 2S albumins are the major allergens involved in severe food allergy to nuts, seeds, and legumes. We aimed to isolate, clone, and express 2S albumin from hazelnut and determine its allergenicity. METHODS: 2S albumin from hazelnut extract was purified using size exclusion chromatography and RP-HPLC. After N-terminal sequencing, degenerated and poly-d(T) primers were used to clone the 2S albumin sequence from hazelnut cDNA. After expression in Escherichia coli and affinity purification, IgE reactivity was evaluated by Immunoblot/ImmunoCAP (inhibition) analyses using sera of nut-allergic patients. RESULTS: N-terminal sequencing of a approximately 10 kDa peak from size exclusion chromatography/RP-HPLC gave two sequences highly homologous to pecan 2S albumin, an 11 amino acid (aa) N-terminal and a 10 aa internal peptide. The obtained clone (441 bp) encoded a 147 aa hazelnut 2S albumin consisting of a putative signal peptide (22 aa), a linker peptide (20 aa), and the mature protein sequence (105 aa). The latter was successfully expressed in E. coli. Both recombinant and natural 2S albumin demonstrated similar IgE reactivity in Immunoblot/ImmunoCAP (inhibition) analyses. CONCLUSION: We confirmed the postulated role of hazelnut 2S albumin as an allergen. The availability of recombinant molecules will allow establishing the importance of hazelnut 2S albumin for hazelnut allergy.
SCOPE: 2S albumins are the major allergens involved in severe food allergy to nuts, seeds, and legumes. We aimed to isolate, clone, and express 2S albumin from hazelnut and determine its allergenicity. METHODS: 2S albumin from hazelnut extract was purified using size exclusion chromatography and RP-HPLC. After N-terminal sequencing, degenerated and poly-d(T) primers were used to clone the 2S albumin sequence from hazelnut cDNA. After expression in Escherichia coli and affinity purification, IgE reactivity was evaluated by Immunoblot/ImmunoCAP (inhibition) analyses using sera of nut-allergicpatients. RESULTS: N-terminal sequencing of a approximately 10 kDa peak from size exclusion chromatography/RP-HPLC gave two sequences highly homologous to pecan 2S albumin, an 11 amino acid (aa) N-terminal and a 10 aa internal peptide. The obtained clone (441 bp) encoded a 147 aa hazelnut 2S albumin consisting of a putative signal peptide (22 aa), a linker peptide (20 aa), and the mature protein sequence (105 aa). The latter was successfully expressed in E. coli. Both recombinant and natural 2S albumin demonstrated similar IgE reactivity in Immunoblot/ImmunoCAP (inhibition) analyses. CONCLUSION: We confirmed the postulated role of hazelnut 2S albumin as an allergen. The availability of recombinant molecules will allow establishing the importance of hazelnut 2S albumin for hazelnut allergy.
Authors: Laurian Zuidmeer-Jongejan; Montserrat Fernández-Rivas; Marcel Gt Winter; Jaap H Akkerdaas; Colin Summers; Ans Lebens; André C Knulst; Piet Schilte; Peter Briza; Gabriele Gadermaier; Ronald van Ree Journal: Clin Transl Allergy Date: 2014-02-02 Impact factor: 5.871
Authors: Sabine Pfeifer; Merima Bublin; Pawel Dubiela; Karin Hummel; Judith Wortmann; Gerhard Hofer; Walter Keller; Christian Radauer; Karin Hoffmann-Sommergruber Journal: Mol Nutr Food Res Date: 2015-08-06 Impact factor: 5.914
Authors: Nuria Prieto; Carmen Burbano; Elisa Iniesto; Julia Rodríguez; Beatriz Cabanillas; Jesus F Crespo; Mercedes M Pedrosa; Mercedes Muzquiz; Juan Carlos Del Pozo; Rosario Linacero; Carmen Cuadrado Journal: Foods Date: 2014-05-05
Authors: Stephen C Dreskin; Stef J Koppelman; Sandra Andorf; Kari C Nadeau; Anjeli Kalra; Werner Braun; Surendra S Negi; Xueni Chen; Catherine H Schein Journal: J Allergy Clin Immunol Date: 2020-11-18 Impact factor: 10.793