Abstract
CdSe/ZnS core/shell quantum dots (QDs) are functionalized with mercaptoundecanoic acid (MUA) and subsequently covered with poly-l-lysine (PLL) as the template for the formation of the silica outer shell. This nanocomposite is used as a transduction and stabilization system for optical biosensor development. The covalent immobilization of the enzyme acetylcholinesterase from Drosophila melanogaster (AChE) during the formation of the biomimetically synthesized silica is used here as a model, relatively unstable enzyme, as a proof of principle. The enzyme is successfully immobilized onto the QDs and then stabilized by the PLL capping and the subsequent formation of the outer nanoporous silica thin shell, giving rise to the QD/AChE/PLL/silica biosensor. It is shown that the poly-l-lysine templated silica outer shell does not modify the optical properties of the quantum dots, while it protects the enzyme from unfolding and denaturation. The small pores of the silica shell allow for the free diffusion of the analyte to the active center of the enzyme, while it does not allow for the proteases to reach the enzyme. The response of the QD/AChE/PLL/silica nano-biosensor to its substrate, acetylcholine chloride, is evaluated by monitoring the changes in the QDs’ photoluminescence which are related to the changes in pH. These pH changes of the surrounding environment of the QDs are induced by the enzymatic reaction, and are associated with the analyte concentration in the solution. The biodetection system proposed is shown to be stable with a storage lifetime of more than 2 months. The data presented provides the grounds for the application of this nanostructured biosensor for the detection of AChE inhibitors.
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Acknowledgments
This work is being supported by the programs NANOMYC (Contract No. 036812) and SANTS (Contract No. 033254) of the European Commission. The authors would like to thank Mrs. Sevasti Papadogiorgaki and Mrs. Alexandra Siakouli from the “Vassilis Galanopoulos” Electron Microscopy Laboratory, for assistance with the transmission electron microscopy.
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Buiculescu, R., Hatzimarinaki, M. & Chaniotakis, N.A. Biosilicated CdSe/ZnS quantum dots as photoluminescent transducers for acetylcholinesterase-based biosensors. Anal Bioanal Chem 398, 3015–3021 (2010). https://doi.org/10.1007/s00216-010-4253-z
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DOI: https://doi.org/10.1007/s00216-010-4253-z