Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system

Optimization of three enzyme immunoassays of very high sensitivity using three antiprolamin monoclonal antibodies (MAbs) (13B4, 11C4 and 12A1) is presented here. These MAbs are specific for those prolamins toxic for coeliac patients, as determined by immunoblotting analysis. Biotinylated MAbs were u...

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Autores principales: Chirdo, Fernando Gabriel, Añón, María Cristina, Fossati, Carlos Alberto
Formato: Articulo
Lenguaje:Inglés
Publicado: 1998
Materias:
Ciencias Exactas
Anti-gliadin monoclonal antibodies
gliadin quantification
ELISA
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/123136
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-123136
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Anti-gliadin monoclonal antibodies
gliadin quantification
ELISA
spellingShingle Ciencias Exactas
Anti-gliadin monoclonal antibodies
gliadin quantification
ELISA
Chirdo, Fernando Gabriel
Añón, María Cristina
Fossati, Carlos Alberto
Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
topic_facet Ciencias Exactas
Anti-gliadin monoclonal antibodies
gliadin quantification
ELISA
description Optimization of three enzyme immunoassays of very high sensitivity using three antiprolamin monoclonal antibodies (MAbs) (13B4, 11C4 and 12A1) is presented here. These MAbs are specific for those prolamins toxic for coeliac patients, as determined by immunoblotting analysis. Biotinylated MAbs were used in two of the assays. In a competitive ELISA, the binding of each biotinylated MAb to a gliadin‐coated solid phase was inhibited by gliadin in the fluid phase. The best result was obtained using the biotinylated MAbl3B4 (detection limit: 20 ng ml⁻¹). With regard to capture ELISA, we tested the performance of the three MAbs. In this sandwich ELISA, the MAb used for antigenic capture was the same as that used as secondary biotinylated antibody. The MAbl2Al had the best performance (detection limit: 1 ng ml⁻¹). The use of biotin‐labelled gliadin in a quantitative immunoassay with a detection limit of 5 ng ml⁻¹ is also reported. This assay involves an antigenic capture using the MAbl2Al followed by a competition between biotinylated and non-biotinylated gliadin. We have found the use of the streptavidin-biotin interaction as signal amplification system to be very useful. This technique, as far as we know, has not been previously reported for gliadin quantification.
format Articulo
Articulo
author Chirdo, Fernando Gabriel
Añón, María Cristina
Fossati, Carlos Alberto
author_facet Chirdo, Fernando Gabriel
Añón, María Cristina
Fossati, Carlos Alberto
author_sort Chirdo, Fernando Gabriel
title Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
title_short Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
title_full Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
title_fullStr Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
title_full_unstemmed Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
title_sort development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system
publishDate 1998
url http://sedici.unlp.edu.ar/handle/10915/123136
work_keys_str_mv AT chirdofernandogabriel developmentofhighsensitiveenzymeimmunoassaysforgliadinquantificationusingthestreptavidinbiotinamplificationsystem
AT anonmariacristina developmentofhighsensitiveenzymeimmunoassaysforgliadinquantificationusingthestreptavidinbiotinamplificationsystem
AT fossaticarlosalberto developmentofhighsensitiveenzymeimmunoassaysforgliadinquantificationusingthestreptavidinbiotinamplificationsystem
bdutipo_str Repositorios
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