An alternative to Western blot analysis using RNA aptamer-functionalized quantum dots

Abstract

To make full use both of optical properties of quantum dots (QDs) and of specific interactions between aptamers and their ligands of interest, we employed QD-conjugated RNA aptamer interactions with histidine tag. QDs offer revolutionary fluorescence performance due to their long-term photostability, brilliant colors, fixability, and narrow, symmetrical emission spectra, and aptamers are known to specifically bind to their target molecules, including metal ions, small molecules, and macromolecules. In this study, we have synthesized RNA aptamer-functionalized QDs, and demonstrated their application to specific protein detection, as an alternative to the conventional Western blot analysis. We observed that our RNA aptamer-functionalized QD system dramatically reduced the time and effort required for conventional Western blot analysis, whereas the selectivity was comparable to that of the conventionally available anti-histidine tag antibody and the sensitivity was comparable to that of the Coomassie blue staining method. In principle, owing to the remarkable optical properties of QDs and a wide versatility of aptamers for selection, our system can harness the high brightness, stability and reusability to quantitatively detect aptamer-recognizable proteins. Furthermore, multiplex detection for several proteins on a single blot can be achieved by our new method, which thus may be able to facilitate and simplify the routinely used protein detection procedure, and make a variety of proteomics analysis possible. (C) 2010 Elsevier Ltd. All rights reserved.