Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1

Research output: Contribution to journalJournal article

Standard

Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1. / Holm, Jens; Ferreras, Mercedes; Ipsen, Henrik; Würtzen, Peter A; Gajhede, Michael; Larsen, Jørgen N; Lund, Kaare; Spangfort, Michael D.

In: Journal of Biological Chemistry, Vol. 286, No. 20, 20.05.2011, p. 17569-17578.

Research output: Contribution to journalJournal article

Harvard

Holm, J, Ferreras, M, Ipsen, H, Würtzen, PA, Gajhede, M, Larsen, JN, Lund, K & Spangfort, MD 2011, 'Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1', Journal of Biological Chemistry, vol. 286, no. 20, pp. 17569-17578. https://doi.org/10.1074/jbc.M110.194878

APA

Holm, J., Ferreras, M., Ipsen, H., Würtzen, P. A., Gajhede, M., Larsen, J. N., ... Spangfort, M. D. (2011). Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1. Journal of Biological Chemistry, 286(20), 17569-17578. https://doi.org/10.1074/jbc.M110.194878

Vancouver

Holm J, Ferreras M, Ipsen H, Würtzen PA, Gajhede M, Larsen JN et al. Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1. Journal of Biological Chemistry. 2011 May 20;286(20):17569-17578. https://doi.org/10.1074/jbc.M110.194878

Author

Holm, Jens ; Ferreras, Mercedes ; Ipsen, Henrik ; Würtzen, Peter A ; Gajhede, Michael ; Larsen, Jørgen N ; Lund, Kaare ; Spangfort, Michael D. / Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 20. pp. 17569-17578.

Bibtex

@article{dab64cb7c49c46499370b5621a3c15b7,
title = "Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1",
abstract = "Birch-allergic patients often experience oral allergy syndrome upon ingestion of vegetables and fruits, most prominently apple, that is caused by antibody cross-reactivity of the IgE antibodies in patients to proteins sharing molecular surface structures with the major birch pollen group 1 allergen from Betula verrucosa (Bet v 1). Still, to what extent two molecular surfaces need to be similar for clinically relevant antibody cross-reactivity to occur is unknown. Here, we describe the grafting of a defined conformational antibody epitope from Bet v 1 onto the surface of the homologous apple allergen Malus domestica group 1 (Mal d 1). Engineering of the epitope was accomplished by genetic engineering substituting amino acid residues in Mal d 1 differing between Bet v 1 and Mal d 1 within the epitope defined by the mAb BV16. The kinetic parameters characterizing the antibody binding interaction to Bet v 1 and to the mutated Mal d 1 variant, respectively, were assessed by Biacore experiments demonstrating indistinguishable binding kinetics. This demonstrates that a conformational epitope defined by a high affinity antibody-allergen interaction can successfully be grafted onto a homologous scaffold molecule without loss of epitope functionality. Furthermore, we show that increasing surface similarity to Bet v 1 of Mal d 1 variants by substitution of 6-8 residues increased the ability to trigger basophil histamine release with blood from birch-allergic patients not responding to natural Mal d 1. Conversely, reducing surface similarity to Bet v 1 of a Mal d 1 variant by substitution of three residues abolished histamine release in one patient reacting to Mal d 1.",
keywords = "Former Faculty of Pharmaceutical Sciences",
author = "Jens Holm and Mercedes Ferreras and Henrik Ipsen and W{\"u}rtzen, {Peter A} and Michael Gajhede and Larsen, {J{\o}rgen N} and Kaare Lund and Spangfort, {Michael D}",
note = "{\circledC} 2011 by The American Society for Biochemistry and Molecular Biology, Inc.",
year = "2011",
month = "5",
day = "20",
doi = "10.1074/jbc.M110.194878",
language = "English",
volume = "286",
pages = "17569--17578",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "20",

}

RIS

TY - JOUR

T1 - Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1

AU - Holm, Jens

AU - Ferreras, Mercedes

AU - Ipsen, Henrik

AU - Würtzen, Peter A

AU - Gajhede, Michael

AU - Larsen, Jørgen N

AU - Lund, Kaare

AU - Spangfort, Michael D

N1 - © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

PY - 2011/5/20

Y1 - 2011/5/20

N2 - Birch-allergic patients often experience oral allergy syndrome upon ingestion of vegetables and fruits, most prominently apple, that is caused by antibody cross-reactivity of the IgE antibodies in patients to proteins sharing molecular surface structures with the major birch pollen group 1 allergen from Betula verrucosa (Bet v 1). Still, to what extent two molecular surfaces need to be similar for clinically relevant antibody cross-reactivity to occur is unknown. Here, we describe the grafting of a defined conformational antibody epitope from Bet v 1 onto the surface of the homologous apple allergen Malus domestica group 1 (Mal d 1). Engineering of the epitope was accomplished by genetic engineering substituting amino acid residues in Mal d 1 differing between Bet v 1 and Mal d 1 within the epitope defined by the mAb BV16. The kinetic parameters characterizing the antibody binding interaction to Bet v 1 and to the mutated Mal d 1 variant, respectively, were assessed by Biacore experiments demonstrating indistinguishable binding kinetics. This demonstrates that a conformational epitope defined by a high affinity antibody-allergen interaction can successfully be grafted onto a homologous scaffold molecule without loss of epitope functionality. Furthermore, we show that increasing surface similarity to Bet v 1 of Mal d 1 variants by substitution of 6-8 residues increased the ability to trigger basophil histamine release with blood from birch-allergic patients not responding to natural Mal d 1. Conversely, reducing surface similarity to Bet v 1 of a Mal d 1 variant by substitution of three residues abolished histamine release in one patient reacting to Mal d 1.

AB - Birch-allergic patients often experience oral allergy syndrome upon ingestion of vegetables and fruits, most prominently apple, that is caused by antibody cross-reactivity of the IgE antibodies in patients to proteins sharing molecular surface structures with the major birch pollen group 1 allergen from Betula verrucosa (Bet v 1). Still, to what extent two molecular surfaces need to be similar for clinically relevant antibody cross-reactivity to occur is unknown. Here, we describe the grafting of a defined conformational antibody epitope from Bet v 1 onto the surface of the homologous apple allergen Malus domestica group 1 (Mal d 1). Engineering of the epitope was accomplished by genetic engineering substituting amino acid residues in Mal d 1 differing between Bet v 1 and Mal d 1 within the epitope defined by the mAb BV16. The kinetic parameters characterizing the antibody binding interaction to Bet v 1 and to the mutated Mal d 1 variant, respectively, were assessed by Biacore experiments demonstrating indistinguishable binding kinetics. This demonstrates that a conformational epitope defined by a high affinity antibody-allergen interaction can successfully be grafted onto a homologous scaffold molecule without loss of epitope functionality. Furthermore, we show that increasing surface similarity to Bet v 1 of Mal d 1 variants by substitution of 6-8 residues increased the ability to trigger basophil histamine release with blood from birch-allergic patients not responding to natural Mal d 1. Conversely, reducing surface similarity to Bet v 1 of a Mal d 1 variant by substitution of three residues abolished histamine release in one patient reacting to Mal d 1.

KW - Former Faculty of Pharmaceutical Sciences

U2 - 10.1074/jbc.M110.194878

DO - 10.1074/jbc.M110.194878

M3 - Journal article

C2 - 21454600

VL - 286

SP - 17569

EP - 17578

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 20

ER -

ID: 34516050