Argentina
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Integrated bio-affinity nano-platform into a microfluidic immunosensor based on monoclonal bispecific trifunctional antibodies for the electrochemical determination of epithelial cancer biomarker
Fecha
2017-01Registro en:
Bravo, Karina; Ortega, Francisco G.; Messina, Germán Alejandro; Sanz Ferramola, Maria Isabel; Fernández Baldo, Martín Alejandro; et al.; Integrated bio-affinity nano-platform into a microfluidic immunosensor based on monoclonal bispecific trifunctional antibodies for the electrochemical determination of epithelial cancer biomarker; Elsevier Science; Clinica Chimica Acta; 464; 1-2017; 64-71
0009-8981
CONICET Digital
CONICET
Autor
Bravo, Karina
Ortega, Francisco G.
Messina, Germán Alejandro
Sanz Ferramola, Maria Isabel
Fernández Baldo, Martín Alejandro
Raba, Julio
Resumen
Background: The epithelial cell adhesion molecule (EpCAM) is a biomarker that is highly overexpressed on the surface of epithelial carcinoma cells. In this study, silver nanoparticles covered with polyvinyl alcohol (AgNPs-PVA) were synthesized, characterized and used in a microfluidic immunosensor based on the use of anti-EpCAM recombinant antibodies as a trapping agent.
Methods: The concentration of trapped EpCAM is then electrochemically quantified by HRP-conjugated anti-EpCAM-antibody. HRP reacted with its enzymatic substrate in a redox process which resulted in the appearance of a current whose magnitude (at a working voltage as low as − 0.10 V) is directly proportional to the concentration of EpCAM.
Results: Under optimized conditions, the detection limits for the microfluidic immunosensor and a commercial ELISA were 0.8 and 13.9 pg/L, respectively. The within-assay and between-assay coefficients of variation are below 6.5% for the proposed method. The immunosensor was validated by analyzing patient samples, and a good correlation with a commercial ELISA was obtained.
Conclusions: The good analytical performance is attributed to the efficient immobilization of the anti-EpCAM recombinant antibodies on the AgNPs-PVA, and its high specificity for EpCAM. This microfluidic immunosensor is intended for use in diagnosis and prognosis of epithelial cancer, to monitor the disease, and to assess therapeutic efficacy.