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  • Overcoming mass transport limitations to achieve femtomolar detection limits on silicon protein microarrays.

Overcoming mass transport limitations to achieve femtomolar detection limits on silicon protein microarrays.

Analytical biochemistry (2011-08-02)
Marina Cretich, Marta Bagnati, Francesco Damin, Laura Sola, Marcella Chiari
ABSTRACT

Here we combine the use of fluorescence-enhancing silicon substrates coated by copoly(DMA-NAS-MAPS), a ter-copolymer based on N,N-dimethylacrylamide (DMA), N-acryloyloxysuccinimide (NAS), and 3-(trimethoxysilyl)propyl-methacrylate (MAPS), with an efficient dynamic incubation to overcome mass transport limitations and obtain femtomolar limits of detection. The high sensitivity was obtained with a conventional microarray scanner without the use of any sophisticated detection strategy or protocol. When the method was applied, an improvement of the analytical sensitivity of approximately three orders of magnitude was achieved for antibody detection when compared with the same assay performed on regular glass slides and static conditions. Moreover, limits of detection of 45 and 54 pg/ml were obtained for hepatitis B superficial antigen and HIV p24 antigen, respectively.

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Sigma-Aldrich
N,N-Dimethylacrylamide, 99%, contains 500 ppm monomethyl ether hydroquinone as inhibitor