Journal
TALANTA
Volume 119, Issue -, Pages 492-497Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2013.11.063
Keywords
Surface plasmon resonance (SPR); Tumor necrosis factor alpha (TNF alpha); Biosensor; Human serum; Gold nanoparticles
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Funding
- Microtecnologias para biodispositivos y deteccion de biomarcadores (Etortek MIBIO2, Basque Government)
- Dispositivos, servicios y aplicaciones para el hospital Inteligente y Extendido (Etorgai OSI+, Basque Government)
- Desarrollo, validacion y aplicabilidad clinica de un nuevo test serico para la deteccion de TNFalpha (Diputacion Foral de Guipuzkoa)
- Instituto de Salud Carlos III
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A simple method for the detection of TNF-alpha protein biomarker in human serum with great sensitivity has been developed using a surface plasmon resonance biosensor. Signal amplification based on a sandwich immunoassay including gold nanoparticles was used. Detection in serum proved to be challenging due to high undesirable non-specific binding to the sensor surface stemming from the matrix nature of the sample. After optimization of the assay parameters and, in the case of serum, of a sample dilution buffer to minimize the non-specific binding, very low limits of detection were achieved: 11.6 pg/mL (211 fM) and 54.4 pg/mL (989 fM) for spiked buffer and human serum respectively. The amplification steps with high affinity biotinylated antibodies and streptavidin-fuctionalized nanoparticles greatly enhanced the signal with the advantage of additional specificity. Due to its simplicity and sensitivity, the immunoassay has proved feasible to be used for detection of low concentration biomarkers in real samples. (C) 2013 Elsevier B.V. All rights reserved.
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