Mimotopes identify conformational epitopes on parvalbumin, the major fish allergen

Untersmayr, Eva; Szalai, Krisztina; Riemer, Angelika B.; Hemmer, Wolfgang; Swoboda, Ines; Hantusch, Brigitte; Schöll, Isabella; Spitzauer, Susanne; Scheiner, Otto; Jarisch, Reinhart; Boltz‐Nitulescu, George; Jensen‐Jarolim, Erika

doi:10.1016/j.molimm.2005.07.038

Cited by 85 publications

(61 citation statements)

References 49 publications

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“…The immunologically reactive sites were located on the junction between the AB and CD domains (AAs 33–44), on the junction between the CD and EF domains (AAs 65–74) and on the calcium-binding loop of the EF domain (AAs 88–96). These results were confirmed in 2006 by Untersmayr et al [31 ]with computational mimotope matching using the three-dimensional model of carp parvalbumin. Furthermore, in this particular study, they suggested that all three regions together may be involved in the antigen-antibody interaction.…”

Section: Discussionsupporting

confidence: 56%

Epitope Mapping of Atlantic Salmon Major Allergen by Peptide Microarray Immunoassay

Pérez‐Gordo

Lin

Bardina

et al. 2011

Int Arch Allergy Immunol

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Background: IgE epitope mapping of allergens reveals important information about antigen elicitors involved in allergic reactions. The peptide-based microarray immunoassay offers an advantage of scale and parallel design over previous methods of epitope mapping. It has been used to map epitopes of some food allergens but has never been used with fish allergens. Objective: We sought to develop a peptide microarray immunoassay to map allergenic fish epitopes of two isoforms of Atlantic salmon (Salmo salar) parvalbumin, Sal s 1 beta 1 and Sal s 1 beta 2. Methods: Sera from 16 fish-allergic patients with specific IgE to salmon parvalbumin were used. Twelve healthy volunteers were used as negative controls. A library of overlapping peptides was synthesized commercially, representing the primary sequence of Sal s 1 beta 1 and Sal s 1 beta 2. Peptides were used to analyze allergen-specific IgE antibodies by immunolabeling with patient sera. Results: Three antigenic regions, not previously described, were identified in Sal s 1 beta 1. Two of them correlated with those previously reported in Gad c 1, parvalbumin from Baltic cod (Gadus callarias). No allergenic regions were found in Sal s 1 beta 2. This could be explained by crucial amino acid substitutions between isoforms. Conclusions: We have identified three antigenic regions in Sal s 1 beta 1 using a peptide microarray immunoassay. These three sequential epitopes formed a unique antigenic determinant in the three-dimensional model of the protein. In addition, we proved that isoforms from the same protein might have a different allergenic behavior.

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Section: Discussionsupporting

confidence: 56%

Epitope Mapping of Atlantic Salmon Major Allergen by Peptide Microarray Immunoassay

Pérez‐Gordo

Lin

Bardina

et al. 2011

Int Arch Allergy Immunol

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“…While Ara h 1 does not need to survive the digestion process as an intact protein or as large fragments to react with the immune system for induction of a specific immune response, this could still be the case for other food allergens. Previous studies examining the influence of digestion on the allergenic potential of other food proteins have revealed digestion to be an effective approach for a significant impairment of sensitization 35,36 and IgE-binding capacity. 35,37,38 We have previously shown that mixtures of peptides smaller than 2−5 kDa, which was generally thought to be the lower size limit for a peptide with inherent sensitizing capacity, 31 may still act as a "complete" allergen, 39 being able to sensitize, elicit allergic reaction, and react with IgE.…”

Section: Journal Of Agricultural and Food Chemistrymentioning

confidence: 99%

Digested Ara h 1 Loses Sensitizing Capacity When Separated into Fractions

Bøgh

Barkholt²,

Rigby

et al. 2012

J. Agric. Food Chem.

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The major peanut allergen Ara h 1 is an easily digestible protein under physiological conditions. The present study revealed that pepsin digestion products of Ara h 1 retained the sensitizing potential in a Brown Norway rat model, while this sensitizing capacity was lost by separating the digest into fractions by gel permeation chromatography. Protein chemical analysis showed that the peptide composition as well as the aggregation profiles of the fractions of Ara h 1 digest differed from that of the whole pool. These results indicate that the sensitizing capacity of digested Ara h 1 is a consequence of the peptides being in an aggregated state resembling the intact molecule or that most peptides of the digests need to be present in the same solution, having a synergistic or adjuvant effect and thereby augmenting the immune response against other peptides.

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“…This approach is not always available to investigators and can be laborious and time consuming. Another approach is to use phage display of random peptides and IgE antibodies from allergic donors, in combination with computational approaches, to determine important conformational epitopes [21,22,23]. However, the phage display approach has not been validated by comparison with the co-crystal structure in a blind test.…”

Section: Introductionmentioning

confidence: 99%

Validation of a Phage Display and Computational Algorithm by Mapping a Conformational Epitope of Bla g 2

Tiwari

Negi

Braun

et al. 2011

Int Arch Allergy Immunol

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Background: Bla g 2, one of the major cockroach allergens, induces a strong IgE response against conformational epitopes, and on reexposure, sensitized individuals often display symptoms of allergic rhinitis and asthma. The aim of the current study was to perform a test of the efficacy of a modified phage display screening, characterization of selected phages and an automated algorithm, EpiSearch, in locating an important conformational epitope. Methods: The monoclonal antibody 7C11, which partially inhibits the binding of patient IgE antibodies to Bla g 2, was used to screen a random peptide phage library. After 3 rounds of panning, 32 phage clones were isolated and the amino acid sequences of their peptides were determined. The relative affinity and specificity of the binding of these peptides to 7C11 were tested in ELISAs. The amino acid composition of these peptides was then matched with clusters of residues on the surface of the 3-dimensional (3D) structure of Bla g 2, using our EpiSearch algorithm. Results: The amino acid sequences of the peptides on selected phages differed at only one position, occupied by 1 of 2 negatively charged residues. The two 12-mer sequences bound to 7C11 with similar avidity and specificity. There was good concordance between the residues in the 3D clusters identified from our phage display/computational method with the co-crystal structural analysis. Conclusion: Conformational epitopes may be mapped through screening of clones from random peptide phage display libraries and EpiSearch.

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Mimotopes identify conformational epitopes on parvalbumin, the major fish allergen

Cited by 85 publications

References 49 publications

Epitope Mapping of Atlantic Salmon Major Allergen by Peptide Microarray Immunoassay

Epitope Mapping of Atlantic Salmon Major Allergen by Peptide Microarray Immunoassay

Digested Ara h 1 Loses Sensitizing Capacity When Separated into Fractions

Validation of a Phage Display and Computational Algorithm by Mapping a Conformational Epitope of Bla g 2

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