Identification of patatin as a novel allergen for children with positive skin prick test responses to raw potato Ulla Seppälä, MSca,b, Harri Alenius,

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Presentation transcript:

Identification of patatin as a novel allergen for children with positive skin prick test responses to raw potato Ulla Seppälä, MSca,b, Harri Alenius, PhDc, Kristiina Turjanmaa, MDd, Timo Reunala, MDd,e, Timo Palosuo, MDb, Nisse Kalkkinen, PhDa Journal of Allergy and Clinical Immunology Volume 103, Issue 1, Pages 165-171 (January 1999) DOI: 10.1016/S0091-6749(99)70541-5 Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 1 Gel filtration and SDS-PAGE analysis of (NH4 )2 SO4 -precipitated potato proteins from the extract. Fractions from gel filtration chromatography were pooled (I to III) and analyzed in SDS-PAGE. Journal of Allergy and Clinical Immunology 1999 103, 165-171DOI: (10.1016/S0091-6749(99)70541-5) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 2 Result of reversed-phase chromatography of purified patatin. Elution of patatin was performed with linear gradient of acetonitrile (3% to 100% in 60 minutes) at a flow rate of 0.2 mL/min. Also shown in the upper right corner is immunoblot analysis of binding of IgE to purified patatin. Coomassie Brilliant Blue–stained patatin (~2 μg/mL) on PVDF-membrane (1) , sera from patient number 7 (2) , and sera from patient number 18 are shown (3) . Journal of Allergy and Clinical Immunology 1999 103, 165-171DOI: (10.1016/S0091-6749(99)70541-5) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 3 Matrix-assisted laser desorption ionization time-of-flight mass spectrum of purified patatin. Journal of Allergy and Clinical Immunology 1999 103, 165-171DOI: (10.1016/S0091-6749(99)70541-5) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 4 Occurrence of IgE-antibodies against purified patatin by ELISA. Microtiterplates were coated with patatin at a concentration of 2.0 μg/ml, and the sera were diluted 1:10. Atopic children (n = 27) with a positive SPT with raw potato (1), control children from the National Vaccination Program (n = 44) (2), and atopic control children (n = 10) (3) are shown. Cut-off level for apositive response (mean OD value + 3 SD of the 44 control subjects) is indicated by horizontal line. Journal of Allergy and Clinical Immunology 1999 103, 165-171DOI: (10.1016/S0091-6749(99)70541-5) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 5 Inhibition of IgE binding to solid-phase total protein extract of potato by different concentrations of purified patatin in ELISA. Patatin inhibited approximately 75% of the IgE binding of pooled serum IgE (diluted 1:20) to the total protein extract. Journal of Allergy and Clinical Immunology 1999 103, 165-171DOI: (10.1016/S0091-6749(99)70541-5) Copyright © 1999 Mosby, Inc. Terms and Conditions