Por favor, use este identificador para citar o enlazar este ítem: https://hdl.handle.net/20.500.12008/50857

Título:

Determination of Microcystins in Fish Tissue by ELISA and MALDI-TOF MS Using a Highly Specific Single Domain Antibody

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Autor:

Badagian, Natalia
Pírez Schirmer, Macarena
Pérez Parada, Andrés
González-Sapienza, Gualberto
Brena, Beatriz M.

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Artículo

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Palabras clave:

Microcystins
ELISA
MALDI-TOF
Fish
Nanobody

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Año de publicación:

2023

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Resumen:

The development of simple, reliable, and cost-effective methods is critically important to study the spatial and temporal variation of microcystins (MCs) in the food chain. Nanobodies (Nbs), antigen binding fragments from camelid antibodies, present valuable features for analytical applications. Their small antigen binding site offers a focused recognition of small analytes, reducing spurious cross-reactivity and matrix effects. A high affinity and broad cross-reactivity anti-MCsNb, from a llama antibody library, was validated in enzyme linked immunosorbent assay (ELISA), and bound to magnetic particles with an internal standard for pre-concentration in quantitativematrix-assisted laser desorption ionization-time of flight mass spectrometry (Nb-QMALDI MS). Both methods are easy and fast; ELISA provides a global result, while Nb-QMALDI MS allows for the quantification of individual congeners and showed excellent performance in the fish muscle extracts. The ELISA assay range was 1.8–29 ng/g and for Nb-QMALDI, it was 0.29–29 ng/g fish ww. Fifty-five fish from a MC-containing dam were analyzed by both methods. The correlation ELISA/sum of the MC congeners by Nb-QMALDI-MS was very high (r Spearman = 0.9645, p < 0.0001). Using ROC curves, ELISA cut-off limits were defined to accurately predict the sum of MCs by Nb-QMALDI-MS (100% sensitivity; ≥89% specificity). Both methods were shown to be simple and efficient for screening MCs in fish muscle to prioritize samples for confirmatory methods.

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Toxins. 15(2) 84, 2023

Financiadores:

Agencia Nacional de Investigación e Innovación (ANII)
Comisión Sectorial de Investigación Científica (CSIC)
Programa de Desarrollo de las Ciencias Básicas (PEDEClBA)

Citación:

BADAGIAN, N., PÍREZ SCHIMER, M., PÉREZ PARADA, A., y otros. Determination of Microcystins in Fish Tissue by ELISA and MALDI-TOF MS Using a Highly Specific Single Domain Antibody. Toxins [en línea] 2023, 15(2) 84. DOI: 10.3390/toxins15020084

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Licencia Creative Commons Atribución (CC - By 4.0)

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