Probing Protein Kinase (CK2) and Alkaline Phosphatase with CdSe/ZnS Quantum Dots

Freeman, Ronit; Finder, Tali; Gill, Ron; Willner, Itamar

doi:10.1021/nl101052f

Cited by 232 publications

(169 citation statements)

References 34 publications

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“…9A). 11 In addition to sensing configurations that use hole accepting metal complexes to quench QD PL, there are examples that use electron accepting molecules, such as quinones, to monitor enzyme activity 199,200 and intracellular pH, 183 or bipyridinium, to monitor receptor-substrate interactions. 180 Initially, Yildiz et al electrostatically adsorbed a bipyridinium dye to the surface of QDs and found that a macrocyclic receptor, cucurbit [7]uril, could disrupt the CT quenching interaction through competitive host-guest interactions.…”

Section: Bioanalysis and Bioimaging With Quantum Dotsmentioning

confidence: 99%

“…183,199,200,202 Freeman et al functionalized QD 620 with peptides containing a phosphotyrosine residue (8 6 2 per QD) to monitor the activity of alkaline phosphatase (ALP). 200 Initially, QDs were unquenched and retained their PL upon hydrolysis of the phosophotyrosine residue by ALP. However, in the presence of a reporter enzyme, tyrosinase, the resultant tyrosine residue is oxidized to a dopaquinone residue that quenched QD PL via CT. Quantification of ALP was demonstrated over the range 0.05-0.5 units in the presence of 25 units of tyrosinase.…”

Section: Bioanalysis and Bioimaging With Quantum Dotsmentioning

confidence: 99%

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Quantum Dots in Bioanalysis: A Review of Applications across Various Platforms for Fluorescence Spectroscopy and Imaging

2013

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Semiconductor quantum dots (QDs) are brightly luminescent nanoparticles that have found numerous applications in bioanalysis and bioimaging. In this review, we highlight recent developments in these areas in the context of specific methods for fluorescence spectroscopy and imaging. Following a primer on the structure, properties, and biofunctionalization of QDs, we describe select examples of how QDs have been used in combination with steady-state or time-resolved spectroscopic techniques to develop a variety of assays, bioprobes, and biosensors that function via changes in QD photoluminescence intensity, polarization, or lifetime. Some special attention is paid to the use of Förster resonance energy transfer-type methods in bioanalysis, including those based on bioluminescence and chemiluminescence. Direct chemiluminescence, electrochemiluminescence, and charge transfer quenching are similarly discussed. We further describe the combination of QDs and flow cytometry, including traditional cellular analyses and spectrally encoded barcode-based assay technologies, before turning our attention to enhanced fluorescence techniques based on photonic crystals or plasmon coupling. Finally, we survey the use of QDs across different platforms for biological fluorescence imaging, including epifluorescence, confocal, and twophoton excitation microscopy; single particle tracking and fluorescence correlation spectroscopy; super-resolution imaging; near-field scanning optical microscopy; and fluorescence lifetime imaging microscopy. In each of the above-mentioned platforms, QDs provide the brightness needed for highly sensitive detection, the photostability needed for tracking dynamic processes, or the multiplexing capacity needed to elucidate complex systems. There is a clear synergy between advances in QD materials and spectroscopy and imaging techniques, as both must be applied in concert to achieve their full potential.

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Section: Bioanalysis and Bioimaging With Quantum Dotsmentioning

confidence: 99%

Section: Bioanalysis and Bioimaging With Quantum Dotsmentioning

confidence: 99%

Quantum Dots in Bioanalysis: A Review of Applications across Various Platforms for Fluorescence Spectroscopy and Imaging

2013

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“…GSH capped QDs were synthesized based on previously reported procedures [27]. The resulting QDs were dissolved in 200 L borate buffer (pH 7.4, 10 mM).…”

Section: Preparation Of Gsh Capped Qdsmentioning

confidence: 99%

Simultaneous detection of assembly and disassembly of multivalent HA tag and anti‐HA antibody in single in‐capillary assay

Wang

Qin

et al. 2016

Electrophoresis

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Herein, we have developed an in-capillary assay for simultaneous detection of the assembly and disassembly of the multivalent HA tag peptide and antibody. HA tag with hexahistidine at C terminus (YPYDVPDYAG4 H6 , termed YPYDH6 ) was conjugated with quantum dots (QDs) by metal-affinity force to form a multivalent HA tag (QD-YPYDH6 ). QD-YPYDH6 and monoclonal anti-HA antibody (anti-HA) were sequentially injected into the capillary. They were mixed and assembled inside the capillary. The reaction products were online discriminated and detected by fluorescence coupled capillary electrophoresis (CE-FL). For the in-capillary assay, the binding efficiency of the multivalent HA tag and antibody on was influenced by the molar ratio and injection time. Such novel assay could even give out the self-assembly kinetic constant of QDs and YPYDH6 as KD of 34.1 μM with n (binding cooperativeness) of 2.2 by Hill equation. More importantly, the simultaneous detection of the assembly and imidazole (Im) induced disassembly of the QD-YPYDH6 -anti-HA complex was achieved in a single in-capillary assay. Our study demonstrated a new method for the online detection of antigen-antibody interactions.

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“…Several fluorescence probes generated from conjugated polyelectrolyte [11][12][13], quantum dots [14][15][16], noble metal nanoclusters [17,18] and organic fluorophores [19][20][21] have been reported for the detection of ALP activities. Despite many achievements from the above-mentioned successful studies, all of them work under one-photon excitation, which requires a rather short excitation wavelength (usually <500 nm).…”

Section: Introductionmentioning

confidence: 99%

A New Two-Photon Ratiometric Fluorescent Probe for Detecting Alkaline Phosphatase in Living Cells

et al. 2016

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Alkaline phosphatase (ALP) is an important diagnostic indicator of many human diseases. To quantitatively track ALP in biosystems, herein, for the first time, we report an efficient two-photon ratiometric fluorescent probe, termed probe 1 and based on classic naphthalene derivatives with a donor-π-acceptor (D-π-A) structure and deprotection of the phosphoric acid moiety by ALP. The presence of ALP causes the cleave of the phosphate group from naphthalene derivatives and the phosphate group changes the ability of the intramolecular charge transfer (ICT) and remarkably alters the probe's photophysical properties, thus an obvious ratiometric signal with an isoemissive point is observed. The fluorescence intensity ratio displayed a linear relationship against the concentration of ALP in the concentration range from 20 to 180 U/L with the limit of detection of 2.3 U/L. Additionally, the probe 1 is further used for fluorescence imaging of ALP in living cells under one-photon excitation (405 nm) or two-photon excitation (720 nm), which showed a high resolution imaging, thus demonstrating its practical application in biological systems.

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Probing Protein Kinase (CK2) and Alkaline Phosphatase with CdSe/ZnS Quantum Dots

Cited by 232 publications

References 34 publications

Quantum Dots in Bioanalysis: A Review of Applications across Various Platforms for Fluorescence Spectroscopy and Imaging

Quantum Dots in Bioanalysis: A Review of Applications across Various Platforms for Fluorescence Spectroscopy and Imaging

Simultaneous detection of assembly and disassembly of multivalent HA tag and anti‐HA antibody in single in‐capillary assay

A New Two-Photon Ratiometric Fluorescent Probe for Detecting Alkaline Phosphatase in Living Cells

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