A chelating dendritic ligand capped quantum dot: preparation, surface passivation, bioconjugation and specific DNA detection.

Herein we report the synthesis of a new chelating dendritic ligand (CDL) and its use in the preparation a compact, stable and water-soluble quantum dot (QD), and further development of specific DNA sensor. The CDL, which contains a chelative dihydrolipoic acid moiety for strong QD surface anchoring and four dendritic carboxylic acid groups, provides a stable, compact and entangled hydrophilic coating around the QD that significantly increases the stability of the resulting water-soluble QD. A CDL-capped CdSe/ZnS core/shell QD (CDL-QD) has stronger fluorescence than that capped by a monodendate single-chain thiol, 3-mercapto-propionic acid (MPA-QD). In addition, the fluorescence of the CDL-QD can be enhanced by 2.5-fold by treatments with Zn2+ or S2- ions, presumably due to effective passivation of the surface defects. This level of fluorescence enhancement obtained for the CDL-QD is much greater than that for the MPA-QD. Further, by coupling a short single-stranded DNA target to the QD via the CDL carboxylic acid group, a functional QD-DNA conjugate that can resist non-specific adsorption and hybridize quickly to its complementary DNA probe has been obtained. This functional QD-DNA conjugate is suitable for specific quantification of short, labelled complementary probes at the low DNA probe:QD copy numbers via a QD-sensitised dye fluorescence resonance energy transfer (FRET) response with 500 pM sensitivity on a conventional fluorimeter.

[1]  Nicholas A. Kotov,et al.  Albumin−CdTe Nanoparticle Bioconjugates: Preparation, Structure, and Interunit Energy Transfer with Antenna Effect , 2001 .

[2]  A. Pron,et al.  Chelating ligands for nanocrystals' surface functionalization. , 2004, Journal of the American Chemical Society.

[3]  Zhivko Zhelev,et al.  Quantum dot-conjugated hybridization probes for preliminary screening of siRNA sequences. , 2005, Journal of the American Chemical Society.

[4]  M. Bawendi,et al.  CdSe nanocrystal based chem-/bio- sensors. , 2007, Chemical Society reviews.

[5]  Masahiko Hara,et al.  Surface State Analysis of Photobrightening in CdSe Nanocrystal Thin Films , 2003 .

[6]  A. Alivisatos Semiconductor Clusters, Nanocrystals, and Quantum Dots , 1996, Science.

[7]  Liming Ying,et al.  Fluorescence resonance energy transfer between a quantum dot donor and a dye acceptor attached to DNA. , 2005, Chemical communications.

[8]  Igor L. Medintz,et al.  Self-assembled nanoscale biosensors based on quantum dot FRET donors , 2003, Nature materials.

[9]  D. Klenerman,et al.  Fabrication of Three‐Dimensional Surface Structures with Highly Fluorescent Quantum Dots by Surface‐Templated Layer‐by‐Layer Assembly , 2005 .

[10]  Hao Yan,et al.  Quantum dot bioconjugation during core-shell synthesis. , 2008, Angewandte Chemie.

[11]  D. Balding,et al.  HLA Sequence Polymorphism and the Origin of Humans , 2006 .

[12]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[13]  Xiaogang Peng,et al.  Stabilization of inorganic nanocrystals by organic dendrons. , 2002, Journal of the American Chemical Society.

[14]  M. Bawendi,et al.  Compact cysteine-coated CdSe(ZnCdS) quantum dots for in vivo applications. , 2007, Journal of the American Chemical Society.

[15]  Igor L. Medintz,et al.  Solution-phase single quantum dot fluorescence resonance energy transfer. , 2006, Journal of the American Chemical Society.

[16]  E. Hall,et al.  A multi-ion particle sensor. , 2007, Chemical communications.

[17]  Igor L. Medintz,et al.  Enhancing the stability and biological functionalities of quantum dots via compact multifunctional ligands. , 2007, Journal of the American Chemical Society.

[18]  M. Bruchez,et al.  Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots , 2003, Nature Biotechnology.

[19]  Vincent Noireaux,et al.  In Vivo Imaging of Quantum Dots Encapsulated in Phospholipid Micelles , 2002, Science.

[20]  U. Krull,et al.  Adsorption and hybridization of oligonucleotides on mercaptoacetic acid-capped CdSe/ZnS quantum dots and quantum dot-oligonucleotide conjugates. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[21]  Hao Yan,et al.  A facile one-step in situ functionalization of quantum dots with preserved photoluminescence for bioconjugation. , 2007, Journal of the American Chemical Society.

[22]  D. Bunka,et al.  Aptamers come of age – at last , 2006, Nature Reviews Microbiology.

[23]  Liming Ying,et al.  Building Three-Dimensional Surface Biological Assemblies on the Nanometer Scale , 2003 .

[24]  M. Bawendi,et al.  Synthesis and characterization of nearly monodisperse CdE (E = sulfur, selenium, tellurium) semiconductor nanocrystallites , 1993 .

[25]  Rebekah Drezek,et al.  Forming biocompatible and nonaggregated nanocrystals in water using amphiphilic polymers. , 2007, Journal of the American Chemical Society.

[26]  S. Nie,et al.  Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules , 2001, Nature Biotechnology.

[27]  J. Ying,et al.  Synthesis and Cell‐Imaging Applications of Glutathione‐Capped CdTe Quantum Dots , 2007 .

[28]  Taekjip Ha,et al.  Single-molecule quantum-dot fluorescence resonance energy transfer. , 2005, Chemphyschem : a European journal of chemical physics and physical chemistry.

[29]  Igor L. Medintz,et al.  Synthesis of compact multidentate ligands to prepare stable hydrophilic quantum dot fluorophores. , 2005, Journal of the American Chemical Society.

[30]  S. Pathak,et al.  Hydroxylated quantum dots as luminescent probes for in situ hybridization. , 2001, Journal of the American Chemical Society.

[31]  Jagjit Nanda,et al.  Effect of the thiol-thiolate equilibrium on the photophysical properties of aqueous CdSe/ZnS nanocrystal quantum dots. , 2005, Journal of the American Chemical Society.

[32]  Juewen Liu,et al.  Functional nucleic acid sensors. , 2009, Chemical reviews.

[33]  Nikolai Gaponik,et al.  THIOL-CAPPING OF CDTE NANOCRYSTALS: AN ALTERNATIVE TO ORGANOMETALLIC SYNTHETIC ROUTES , 2002 .

[34]  U. Krull,et al.  Towards multi-colour strategies for the detection of oligonucleotide hybridization using quantum dots as energy donors in fluorescence resonance energy transfer (FRET). , 2007, Analytica chimica acta.

[35]  Igor L. Medintz,et al.  A fluorescence resonance energy transfer-derived structure of a quantum dot-protein bioconjugate nanoassembly. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[36]  R. Leblanc,et al.  Peptide-coated CdS quantum dots for the optical detection of copper(II) and silver(I). , 2003, Chemical communications.

[37]  Z. Rosenzweig,et al.  Luminescent CdS quantum dots as selective ion probes. , 2002, Analytical chemistry.

[38]  Dejian Zhou,et al.  Label-free detection of DNA hybridization at the nanoscale: a highly sensitive and selective approach using atomic-force microscopy. , 2003, Angewandte Chemie.

[39]  J. Vela,et al.  "Giant" multishell CdSe nanocrystal quantum dots with suppressed blinking. , 2008, Journal of the American Chemical Society.

[40]  D. Klenerman,et al.  A compact functional quantum Dot-DNA conjugate: preparation, hybridization, and specific label-free DNA detection. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[41]  H. Yeh,et al.  Single-quantum-dot-based DNA nanosensor , 2005, Nature materials.

[42]  Shimon Weiss,et al.  Synthesis and Properties of Biocompatible Water-Soluble Silica-Coated CdSe/ZnS Semiconductor Quantum Dots† , 2001 .

[43]  Peter G Stockley,et al.  Development of smart nanoparticle-aptamer sensing technology. , 2011, Faraday discussions.

[44]  Zeev Rosenzweig,et al.  Synthesis and application of quantum dots FRET-based protease sensors. , 2006, Journal of the American Chemical Society.

[45]  S. Nie,et al.  Quantum dot bioconjugates for ultrasensitive nonisotopic detection. , 1998, Science.

[46]  C. Soeller,et al.  DNA hybridization detection with blue luminescent quantum dots and dye-labeled single-stranded DNA. , 2007, Journal of the American Chemical Society.

[47]  Xiaogang Peng,et al.  Formation of high-quality CdTe, CdSe, and CdS nanocrystals using CdO as precursor. , 2001, Journal of the American Chemical Society.

[48]  Igor L. Medintz,et al.  Fluorescence resonance energy transfer between quantum dot donors and dye-labeled protein acceptors. , 2003, Journal of the American Chemical Society.

[49]  Junghan Lee,et al.  Positively charged compact quantum Dot-DNA complexes for detection of nucleic acids. , 2009, Chemphyschem : a European journal of chemical physics and physical chemistry.

[50]  M. Osborne,et al.  Photodynamics of a single quantum dot: fluorescence activation, enhancement, intermittency, and decay. , 2007, Journal of the American Chemical Society.

[51]  P. Alivisatos The use of nanocrystals in biological detection , 2004, Nature Biotechnology.

[52]  S. Deo,et al.  Rapid, single-step nucleic acid detection , 2008, Analytical and bioanalytical chemistry.

[53]  L. Pu,et al.  Surface treatment to enhance the quantum efficiency of semiconductor nanocrystals , 2004 .

[54]  R. Zentel,et al.  CdSe/ZnS nanocrystals with dye-functionalized polymer ligands containing many anchor groups. , 2005, Angewandte Chemie.

[55]  S. Balasubramanian,et al.  A reversible pH-driven DNA nanoswitch array. , 2006, Journal of the American Chemical Society.

[56]  H. Mattoussi,et al.  Multifunctional ligands based on dihydrolipoic acid and polyethylene glycol to promote biocompatibility of quantum dots , 2009, Nature Protocols.

[57]  E. Doris,et al.  A versatile strategy for quantum dot ligand exchange. , 2007, Journal of the American Chemical Society.

[58]  Eunkeu Oh,et al.  Inhibition assay of biomolecules based on fluorescence resonance energy transfer (FRET) between quantum dots and gold nanoparticles. , 2005, Journal of the American Chemical Society.

[59]  M. Levy,et al.  Quantum‐Dot Aptamer Beacons for the Detection of Proteins , 2005, Chembiochem : a European journal of chemical biology.

[60]  S. Wuister,et al.  Luminescence temperature antiquenching of water-soluble CdTe quantum dots: role of the solvent. , 2004, Journal of the American Chemical Society.

[61]  Warren C. W. Chan,et al.  Quantum Dots in Biological and Biomedical Research: Recent Progress and Present Challenges , 2006 .