Quantification of autoantibodies to annexin V in plasma by an "in house" sandwich ELISA

• Autor(a) principal: Pelusa,Héctor F.
• Data de Publicação: 2012
• Outros Autores: Valdés,María de los Ángeles, Daniele,Stella M., Almará,Adriana M., Svetaz,María J., Bearzotti,Mariela, Paparella,Cecilia V., Bouvet,Beatriz R., Ghersevich,Sergio A., Arriaga,Sandra M.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Journal of the Brazilian Chemical Society (Online)
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532012000300011
• topic: ELISA; annexin V; antibodies; anti-phospholipid syndrome; foetal losses

An "in house" sandwich ELISA for the quantification of plasma anti-annexin V antibodies was developed and validated. The validation parameters studied were: (i) precision, expressed either as the intra- or the inter-assay coefficient of variation (CV), (ii) exactitude, expressed as the percentage deviation between the obtained value and the real value, (iii) limit of detection (LOD), evaluated from the reagents blank and (iv) robustness, obtained by deliberately introducing slight variations in different parameters. Also, a comparison between the "in house" technique and a commercial method was performed. The research revealed that both CV were < 20%, exactitude was within the 100 ± 20% range, limit of detection was below 1 U mL-1 and that slight variations in the technique did not produce any significant variations in the results. Comparison with the commercial method showed an acceptable correlation. It was concluded that the method developed here satisfactorily accomplishes the parameters of standardization and validation for an immunoassay.