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Philippe A. Eigenmann, MD,a, A. Wesley Burks, MD,b, Gary A

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Presentation on theme: "Philippe A. Eigenmann, MD,a, A. Wesley Burks, MD,b, Gary A"— Presentation transcript:

1 Identification of unique peanut and soy allergens in sera adsorbed with cross-reacting antibodies 
Philippe A. Eigenmann, MD,a, A.Wesley Burks, MD,b, Gary A. Bannon, PhD,b, Hugh A. Sampson, MDa  Journal of Allergy and Clinical Immunology  Volume 98, Issue 5, Pages (November 1996) DOI: /S (96) Copyright © 1996 Mosby, Inc. Terms and Conditions

2 FIG. 1 Results of IgE binding to soy and peanut by ELISA from nonabsorbed sera and after successive passes over the soy-affinity chromatography column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient BM. C, Patient DH. D, Patient AT. E, Patient DT. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

3 FIG. 1 Results of IgE binding to soy and peanut by ELISA from nonabsorbed sera and after successive passes over the soy-affinity chromatography column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient BM. C, Patient DH. D, Patient AT. E, Patient DT. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

4 FIG. 1 Results of IgE binding to soy and peanut by ELISA from nonabsorbed sera and after successive passes over the soy-affinity chromatography column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient BM. C, Patient DH. D, Patient AT. E, Patient DT. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

5 FIG. 1 Results of IgE binding to soy and peanut by ELISA from nonabsorbed sera and after successive passes over the soy-affinity chromatography column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient BM. C, Patient DH. D, Patient AT. E, Patient DT. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

6 FIG. 1 Results of IgE binding to soy and peanut by ELISA from nonabsorbed sera and after successive passes over the soy-affinity chromatography column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient BM. C, Patient DH. D, Patient AT. E, Patient DT. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

7 FIG. 2 IgE antibody binding to whole peanut extract, purified Ara h 1, and Ara h 2 by ELISA before and after soy-specific antibody adsorption over the soy-affinity chromatography column. Each square represents optical density (O.D.) readings for a specific patient. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

8 FIG. 3 Tricine–SDS polyacrylamide gel. Molecular weight marker (MWM), soy, and peanut fractions are stained with amido black 0.1%. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

9 FIG. 4 IgE binding to peanut fractions isolated on tricine–SDS polyacrylamide immunoblots for two patients allergic to peanut and soy (BP and BM), and three soy-tolerant patients with peanut allergy (DH, AT, and DT). The first lane for each patient was reacted with nonadsorbed serum, and the second lane was reacted with soy protein–adsorbed serum. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

10 FIG. 5 Glycine–SDS-PAGE of soy and peanut. The first three lanes represent molecular weight marker (MWM), soy, and peanut stained with amido black 0.1%. The next three sets of lanes show IgE antibody binding to soy and peanut protein fractions with nonadsorbed serum from patient BP, serum passed twice over the soy-affinity chromatography column, and serum adsorbed five times over the column. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

11 FIG. 6 IgE antibody from sera of two patients allergic to peanut and soy (BP and BM) and three soy-tolerant patients with peanut allergy (DH, AT, and DT) binding to tricine–SDS polyacrylamide soy immunoblots. The first lane for each patient was reacted with nonadsorbed serum; the second lane was blotted with soy-adsorbed serum. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

12 FIG. 7 Control immunoblots of soy (s) and peanut (p). A Lanes developed without serum. B, Lanes assayed with serum from a patient lacking antibody to both soy and peanut. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

13 FIG. 8 Relative quantities of IgE binding to whole soy and peanut proteins by ELISA after successive passes over the peanut-affinity column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient DT. OD, Optical density. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

14 FIG. 8 Relative quantities of IgE binding to whole soy and peanut proteins by ELISA after successive passes over the peanut-affinity column. Dotted line represents IgE binding to peanut antigen; solid line represents IgE binding to soy antigen. A, Patient BP. B, Patient DT. OD, Optical density. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

15 FIG. 9 IgE binding to tricine–SDS polyacrylamide peanut and soy immunoblots for one patient allergic to peanut and soy (BP) and one soy-tolerant patient with peanut allergy (DT). The first lane for each patient was reacted with nonadsorbed serum; the second lane was reacted with peanut-adsorbed serum. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

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