Data di Pubblicazione:
2013
Citazione:
Multispot, label-free biodetection at a phantom plastic–water interface / F. Giavazzi, M. Salina, R. Cerbino, M. Bassi, D. Prosperi, E. Ceccarello, F. Damin, L. Sola, M. Rusnati, M. Chiari, B. Chini, T. Bellini, M. Buscaglia. - In: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. - ISSN 0027-8424. - 110:23(2013 Jun 04), pp. 9350-9355. [10.1073/pnas.1214589110]
Abstract:
Recognizing and quantifying specific biomolecules in aqueous
samples are constantly needed in research and diagnostic laboratories.
As the typical detection procedures are rather lengthy and
involve the use of labeled secondary antibodies or other agents to
provide a signal, efforts have been made over the last 10 y to
develop alternative label-free methods that enable direct detection.
We propose and demonstrate an extremely simple, lowcost,
label-free biodetector based on measuring the intensity of
light reflected by the interface between a fluid sample and an
amorphous fluoropolymer substrate having a refractive index very
close to that of water and hosting various antibodies immobilized
in spots. Under these index-matching conditions, the amount of
light reflected by the interface allows straightforward quantification
of the amount of antigen binding to each spot. Using
antibodies targeting heterologous immunoglobulins and antigens
commonly used as markers for diagnoses of hepatitis B and HIV,
we demonstrate the limit of detection of a few picograms per
square millimeter of surface-bound molecules. We also show that
direct and real-time access to the amount of binding molecules
allows the precise extrapolation of adhesion rates, from which the
concentrations of antigens in solution can be estimated down to
fractions of nanograms per milliliter.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
immunoassay; optical biosensor; protein microarray; biomolecular detection; reflective phantom interface
Elenco autori:
F. Giavazzi, M. Salina, R. Cerbino, M. Bassi, D. Prosperi, E. Ceccarello, F. Damin, L. Sola, M. Rusnati, M. Chiari, B. Chini, T. Bellini, M. Buscaglia
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