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

Anal Biochem. 2011 Nov 1;418(1):164-6. doi: 10.1016/j.ab.2011.07.004. Epub 2011 Jul 14.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamides / chemistry
  • HIV Core Protein p24 / analysis
  • HIV Core Protein p24 / chemistry
  • Limit of Detection
  • Methacrylates / chemistry
  • Organosilicon Compounds / chemistry
  • Protein Array Analysis / methods*
  • Sensitivity and Specificity
  • Silicon / chemistry*

Substances

  • Acrylamides
  • HIV Core Protein p24
  • Methacrylates
  • Organosilicon Compounds
  • N,N-dimethylacrylamide
  • 3-(trimethoxysilyl)propyl methacrylate
  • Silicon