Validation of Immunoassay-Based Tools for the Comprehensive Quantification of Aß40 and Aß42 Peptides in Plasma
Pérez-Grijalba, V. ; Fandos, N. ; Canudas, J. ; Insua, D. ; Casabona, D. ; Lacosta, A. M. ; Montañés, M. ; Pesini, P. ; Sarasa Barrio, José Manuel (Universidad de Zaragoza)
Resumen: Recent advances in neuroimaging and cerebrospinal fluid (CSF) biomarker assays have provided evidence of a long preclinical stage of Alzheimer''s disease (AD). This period is being increasingly targeted for secondary prevention trials of new therapies. In this context, the interest of a noninvasive, cost-effective amyloid-ß (Aß) blood-based test does not need to be overstated. Nevertheless, a thorough validation of these bioanalytical methods should be performed as a prerequisite for confident interpretation of clinical results. The aim of this study was to validate ELISA sandwich colorimetric ABtest40 and ABtest42 for the quantification of Aß40 and Aß42 in human plasma. The validation parameters assessed included precision, accuracy, sensitivity, specificity, recovery, and dilution linearity. ABtest40 and ABtest42 proved to be specific for their target peptide using Aß peptides with sequence similar to the target. Mean relative error in the quantification was found to be below 7.5 for both assays, with high intra-assay, inter-assay, and inter-batch precision (CV <9.0 on average). Sensitivity was assessed by determination of the limit of quantification fulfilling precision and accuracy criteria; it was established at 7.60 pg/ml and 3.60 pg/ml for ABtest40 and ABtest42, respectively. Plasma dilution linearity was demonstrated in PBS; however, dilution in a proprietary formulated buffer significantly increased the recovery of both Aß40 and Aß42 masked by matrix interactions, allowing a more comprehensive assessment of the free and total peptide levels in the plasma. In conclusion, both assays were successfully validated as tools for the quantification Aß40 and Aß42 in plasma.
Idioma: Inglés
DOI: 10.3233/JAD-160325
Año: 2016
Publicado en: JOURNAL OF ALZHEIMERS DISEASE 54, 2 (2016), 751-762
ISSN: 1387-2877
Tipo y forma: Article (Published version)
Área (Departamento): Anatomía y Anatomía Patológica Comparadas (Departamento de Anatomía, Embriología y Genética Animal)
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Exportado de SIDERAL (2016-10-19-11:24:53)
Idioma: Inglés
DOI: 10.3233/JAD-160325
Año: 2016
Publicado en: JOURNAL OF ALZHEIMERS DISEASE 54, 2 (2016), 751-762
ISSN: 1387-2877
Tipo y forma: Article (Published version)
Área (Departamento): Anatomía y Anatomía Patológica Comparadas (Departamento de Anatomía, Embriología y Genética Animal)
0
Exportado de SIDERAL (2016-10-19-11:24:53)
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articulos > articulos-por-area > anatomia_y_anatomia_patologica_comparadas
Notice créée le 2016-10-19, modifiée le 2016-10-20
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