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An antibody-based ELISA for quantification of Ani s 1, a major allergen from Anisakis simplex

Published online by Cambridge University Press:  01 April 2008

M. C. ARILLA ,
I. IBARROLA ,
A. MARTÍNEZ ,
J. MONTESEIRÍN ,
J. CONDE  and
J. A. ASTURIAS
M. C. ARILLA
Affiliation:
Research & Development Department, Bial-Arístegui, Alameda Urquijo, 27, 48008 Bilbao, Spain
I. IBARROLA
Affiliation:
Research & Development Department, Bial-Arístegui, Alameda Urquijo, 27, 48008 Bilbao, Spain
A. MARTÍNEZ
Affiliation:
Research & Development Department, Bial-Arístegui, Alameda Urquijo, 27, 48008 Bilbao, Spain
J. MONTESEIRÍN
Affiliation:
Department of Medicine, Immunology and Allergy Service, Virgen Macarena University Hospital, Sevilla Sagrado Corazón Clinic, Sevilla, Spain
J. CONDE
Affiliation:
Sagrado Corazón Clinic, Sevilla, Spain
J. A. ASTURIAS*
Affiliation:
Research & Development Department, Bial-Arístegui, Alameda Urquijo, 27, 48008 Bilbao, Spain
*
*Corresponding author: Bial-Arístegui, R&D; Alameda Urquijo, 27; 48008-Bilbao, Spain. Tel: +34 944 438 000. Fax: +34 944 438 016. E-mail: juan.asturias@bial.com
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Summary

Anisakis simplex is a nematode parasite that can infect humans who have eaten raw or undercooked seafood. Larvae invading the gastrointestinal mucosa excrete/secrete proteins that are implicated in the pathogenesis of anisakiasis and can induce IgE-mediated symptoms. Since Ani s 1 is a potent secreted allergen with important clinical relevance, its measurement could assess the quality of allergenic products used in diagnosis/immunotherapy of Anisakis allergy and track the presence of A. simplex parasites in fish foodstuffs. An antibody-based ELISA for quantification of Ani s 1 has been developed based on monoclonal antibody 4F2 as capture antibody and biotin-labelled polyclonal antibodies against Ani s 1 as detection reagent. The dose-response standard curves, obtained with natural and recombinant antigens, ranged from 4 to 2000 ng/ml and were identical and parallel to that of the A. simplex extract. The linear portion of the dose-response curve with nAni s 1 was between 15 and 250 ng/ml with inter-assay and intra-assays coefficients of variation less than 20% and 10%, respectively. The assay was specific since there was no cross-reaction with other extracts (except Ascaris extracts) and was highly sensitive (detection limit of 1·8 ng/ml), being able to detect Ani s 1 in fish extracts from codfish and monkfish.

Keywords

Anisakis simplexAnisakis allergyAni s 1quantificationmonoclonal antibodyimmunoassayfood allergyrecombinant antigenparasite detection
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 6 , May 2008 , pp. 735 - 740
Copyright
Copyright © 2008 Cambridge University Press

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