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Title: Azide–Alkyne Click Conjugation on Quantum Dots by Selective Copper Coordination

Abstract

Functionalization of nanocrystals is essential for their practical application, but synthesis on nanocrystal surfaces is limited by incompatibilities with certain key reagents. The copper-catalyzed azide-alkyne cycloaddition is among the most useful methods for ligating molecules to surfaces, but has been largely useless for semiconductor quantum dots (QDs) because Cu+ions quickly and irreversibly quench QD fluorescence. To discover nonquenching synthetic conditions for Cu-catalyzed click reactions on QD surfaces, we developed a combinatorial fluorescence assay to screen >2000 reaction conditions to maximize cycloaddition efficiency while minimizing QD quenching. We identify conditions for complete coupling without significant quenching, which are compatible with common QD polymer surfaces and various azide/alkyne pairs. Based on insight from the combinatorial screen and mechanistic studies of Cu coordination and quenching, we find that superstoichiometric concentrations of Cu can promote full coupling if accompanied by ligands that selectively compete with the Cu from the QD surface but allow it to remain catalytically active. Applied to the conjugation of a K+ channel-specific peptidyl toxin to CdSe/ZnS QDs, we synthesize unquenched QD conjugates and image their specific and voltage-dependent affinity for K+ channels in live cells.

Authors:
 [1];  [1];  [2]; ORCiD logo [3]; ORCiD logo [1]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). The Molecular Foundry
  2. Univ. of California, Davis, CA (United States). Dept. of Physiology and Membrane Biology
  3. Univ. of California, Davis, CA (United States). Dept. of Physiology and Membrane Biology
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)
OSTI Identifier:
1460332
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
ACS Nano
Additional Journal Information:
Journal Volume: 12; Journal Issue: 5; Journal ID: ISSN 1936-0851
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
77 NANOSCIENCE AND NANOTECHNOLOGY; quantum dot; CuAAC; quenching; copper; synthesis; combinatorial nanoscience; bioconjugation; high-throughput screen

Citation Formats

Mann, Victor R., Powers, Alexander S., Tilley, Drew C., Sack, Jon T., and Cohen, Bruce E.. Azide–Alkyne Click Conjugation on Quantum Dots by Selective Copper Coordination. United States: N. p., 2018. Web. https://doi.org/10.1021/acsnano.8b00575.
Mann, Victor R., Powers, Alexander S., Tilley, Drew C., Sack, Jon T., & Cohen, Bruce E.. Azide–Alkyne Click Conjugation on Quantum Dots by Selective Copper Coordination. United States. https://doi.org/10.1021/acsnano.8b00575
Mann, Victor R., Powers, Alexander S., Tilley, Drew C., Sack, Jon T., and Cohen, Bruce E.. Mon . "Azide–Alkyne Click Conjugation on Quantum Dots by Selective Copper Coordination". United States. https://doi.org/10.1021/acsnano.8b00575. https://www.osti.gov/servlets/purl/1460332.
@article{osti_1460332,
title = {Azide–Alkyne Click Conjugation on Quantum Dots by Selective Copper Coordination},
author = {Mann, Victor R. and Powers, Alexander S. and Tilley, Drew C. and Sack, Jon T. and Cohen, Bruce E.},
abstractNote = {Functionalization of nanocrystals is essential for their practical application, but synthesis on nanocrystal surfaces is limited by incompatibilities with certain key reagents. The copper-catalyzed azide-alkyne cycloaddition is among the most useful methods for ligating molecules to surfaces, but has been largely useless for semiconductor quantum dots (QDs) because Cu+ions quickly and irreversibly quench QD fluorescence. To discover nonquenching synthetic conditions for Cu-catalyzed click reactions on QD surfaces, we developed a combinatorial fluorescence assay to screen >2000 reaction conditions to maximize cycloaddition efficiency while minimizing QD quenching. We identify conditions for complete coupling without significant quenching, which are compatible with common QD polymer surfaces and various azide/alkyne pairs. Based on insight from the combinatorial screen and mechanistic studies of Cu coordination and quenching, we find that superstoichiometric concentrations of Cu can promote full coupling if accompanied by ligands that selectively compete with the Cu from the QD surface but allow it to remain catalytically active. Applied to the conjugation of a K+ channel-specific peptidyl toxin to CdSe/ZnS QDs, we synthesize unquenched QD conjugates and image their specific and voltage-dependent affinity for K+ channels in live cells.},
doi = {10.1021/acsnano.8b00575},
journal = {ACS Nano},
number = 5,
volume = 12,
place = {United States},
year = {2018},
month = {4}
}

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Works referenced in this record:

Quantum dot bioconjugates for imaging, labelling and sensing
journal, June 2005

  • Medintz, Igor L.; Uyeda, H. Tetsuo; Goldman, Ellen R.
  • Nature Materials, Vol. 4, Issue 6, p. 435-446
  • DOI: 10.1038/nmat1390

Compact high-quality CdSe–CdS core–shell nanocrystals with narrow emission linewidths and suppressed blinking
journal, February 2013

  • Chen, Ou; Zhao, Jing; Chauhan, Vikash P.
  • Nature Materials, Vol. 12, Issue 5
  • DOI: 10.1038/nmat3539

The surface science of nanocrystals
journal, January 2016

  • Boles, Michael A.; Ling, Daishun; Hyeon, Taeghwan
  • Nature Materials, Vol. 15, Issue 2
  • DOI: 10.1038/nmat4526

Energy Transfer with Semiconductor Quantum Dot Bioconjugates: A Versatile Platform for Biosensing, Energy Harvesting, and Other Developing Applications
journal, March 2016


Quantum Dot Surface Chemistry and Functionalization for Cell Targeting and Imaging
journal, March 2015


Fluorescence Resonance Energy Transfer Between Quantum Dot Donors and Dye-Labeled Protein Acceptors
journal, January 2004

  • Clapp, Aaron R.; Medintz, Igor L.; Mauro, J. Matthew
  • Journal of the American Chemical Society, Vol. 126, Issue 1
  • DOI: 10.1021/ja037088b

Development of a Bioorthogonal and Highly Efficient Conjugation Method for Quantum Dots Using Tetrazine−Norbornene Cycloaddition
journal, June 2010

  • Han, Hee-Sun; Devaraj, Neal K.; Lee, Jungmin
  • Journal of the American Chemical Society, Vol. 132, Issue 23
  • DOI: 10.1021/ja101677r

Avidin:  A Natural Bridge for Quantum Dot-Antibody Conjugates
journal, May 2002

  • Goldman, Ellen R.; Balighian, Eric D.; Mattoussi, Hedi
  • Journal of the American Chemical Society, Vol. 124, Issue 22
  • DOI: 10.1021/ja0125570

DNA-Functionalized Quantum Dots: Fabrication, Structural, and Physicochemical Properties
journal, May 2013


Poly(ethylene glycol) Carbodiimide Coupling Reagents for the Biological and Chemical Functionalization of Water-Soluble Nanoparticles
journal, March 2009

  • Shen, Hongyan; Jawaid, Ali M.; Snee, Preston T.
  • ACS Nano, Vol. 3, Issue 4
  • DOI: 10.1021/nn800870r

A Novel Fluorescent Probe for Copper Ions Based on Polymer-modified CdSe/CdS Core/Shell Quantum Dots
journal, January 2011


Tarantula toxins use common surfaces for interacting with Kv and ASIC ion channels
journal, May 2015


Chemoselective tarantula toxins report voltage activation of wild-type ion channels in live cells
journal, October 2014

  • Tilley, D. C.; Eum, K. S.; Fletcher-Taylor, S.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 44
  • DOI: 10.1073/pnas.1406876111

Analysis and Optimization of Copper-Catalyzed Azide–Alkyne Cycloaddition for Bioconjugation
journal, December 2009

  • Hong, Vu; Presolski, Stanislav I.; Ma, Celia
  • Angewandte Chemie International Edition, Vol. 48, Issue 52
  • DOI: 10.1002/anie.200905087

Reliable and Efficient Procedures for the Conjugation of Biomolecules through Huisgen Azide-Alkyne Cycloadditions
journal, August 2011

  • Lallana, Enrique; Riguera, Ricardo; Fernandez-Megia, Eduardo
  • Angewandte Chemie International Edition, Vol. 50, Issue 38
  • DOI: 10.1002/anie.201101019

Fast, Cell-Compatible Click Chemistry with Copper-Chelating Azides for Biomolecular Labeling
journal, May 2012

  • Uttamapinant, Chayasith; Tangpeerachaikul, Anupong; Grecian, Scott
  • Angewandte Chemie International Edition, Vol. 51, Issue 24
  • DOI: 10.1002/anie.201108181

Toward the Syntheses of Universal Ligands for Metal Oxide Surfaces:  Controlling Surface Functionality through Click Chemistry
journal, September 2006

  • White, Meghann A.; Johnson, Jeremiah A.; Koberstein, Jeffrey T.
  • Journal of the American Chemical Society, Vol. 128, Issue 35
  • DOI: 10.1021/ja064041s

How to very efficiently functionalize gold nanoparticles by “click” chemistry
journal, January 2008

  • Boisselier, Elodie; Salmon, Lionel; Ruiz, Jaime
  • Chemical Communications, Issue 44
  • DOI: 10.1039/b812249k

A Stepwise Huisgen Cycloaddition Process: Copper(I)-Catalyzed Regioselective “Ligation” of Azides and Terminal Alkynes
journal, July 2002


Advancements in the mechanistic understanding of the copper-catalyzed azide–alkyne cycloaddition
journal, January 2013

  • Berg, Regina; Straub, Bernd F.
  • Beilstein Journal of Organic Chemistry, Vol. 9
  • DOI: 10.3762/bjoc.9.308

Luminescent detection of Cu(II) ions in aqueous solution using CdSe and CdSe-ZnS quantum dots functionalised with mercaptosuccinic acid
journal, April 2009


Luminescence of Polyethylene Glycol Coated CdSeTe/ZnS and InP/ZnS Nanoparticles in the Presence of Copper Cations
journal, June 2011

  • Beaune, Grégory; Tamang, Sudarsan; Bernardin, Aude
  • ChemPhysChem, Vol. 12, Issue 12
  • DOI: 10.1002/cphc.201100266

Investigations of ion transport through nanoscale polymer membranes by fluorescence quenching of CdSe/CdS quantum dot/quantum rods
journal, January 2016

  • Merkl, Jan-Philip; Wolter, Christopher; Flessau, Sandra
  • Nanoscale, Vol. 8, Issue 14
  • DOI: 10.1039/C5NR08318D

Controlling the Physical and Biological Properties of Highly Fluorescent Aqueous Quantum Dots Using Block Copolymers of Different Size and Shape
journal, September 2013

  • Ostermann, Johannes; Merkl, Jan-Philip; Flessau, Sandra
  • ACS Nano, Vol. 7, Issue 10
  • DOI: 10.1021/nn4037859

Sequential Cation Exchange in Nanocrystals: Preservation of Crystal Phase and Formation of Metastable Phases
journal, November 2011

  • Li, Hongbo; Zanella, Marco; Genovese, Alessandro
  • Nano Letters, Vol. 11, Issue 11
  • DOI: 10.1021/nl202927a

Cu-free click cycloaddition reactions in chemical biology
journal, January 2010

  • Jewett, John C.; Bertozzi, Carolyn R.
  • Chemical Society Reviews, Vol. 39, Issue 4
  • DOI: 10.1039/b901970g

A Strain-Promoted [3 + 2] Azide−Alkyne Cycloaddition for Covalent Modification of Biomolecules in Living Systems
journal, November 2004

  • Agard, Nicholas J.; Prescher, Jennifer A.; Bertozzi, Carolyn R.
  • Journal of the American Chemical Society, Vol. 126, Issue 46
  • DOI: 10.1021/ja044996f

Hydrophobic Nanocrystals Coated with an Amphiphilic Polymer Shell:  A General Route to Water Soluble Nanocrystals
journal, April 2004

  • Pellegrino, Teresa; Manna, Liberato; Kudera, Stefan
  • Nano Letters, Vol. 4, Issue 4, p. 703-707
  • DOI: 10.1021/nl035172j

Dual-Emitting Quantum Dot/Quantum Rod-Based Nanothermometers with Enhanced Response and Sensitivity in Live Cells
journal, June 2012

  • Albers, Aaron E.; Chan, Emory M.; McBride, Patrick M.
  • Journal of the American Chemical Society, Vol. 134, Issue 23
  • DOI: 10.1021/ja302290e

Covalent Protein Labeling and Improved Single-Molecule Optical Properties of Aqueous CdSe/CdS Quantum Dots
journal, June 2017


Tailored Ligand Acceleration of the Cu-Catalyzed Azide−Alkyne Cycloaddition Reaction: Practical and Mechanistic Implications
journal, October 2010

  • Presolski, Stanislav I.; Hong, Vu; Cho, So-Hye
  • Journal of the American Chemical Society, Vol. 132, Issue 41
  • DOI: 10.1021/ja105743g

Sulfated Ligands for the Copper(I)-Catalyzed Azide-Alkyne Cycloaddition
journal, September 2011

  • Wang, Wei; Hong, Senglian; Tran, Andrew
  • Chemistry - An Asian Journal, Vol. 6, Issue 10
  • DOI: 10.1002/asia.201100385

Ligand-Accelerated Cu-Catalyzed Azide−Alkyne Cycloaddition:  A Mechanistic Report
journal, October 2007

  • Rodionov, Valentin O.; Presolski, Stanislav I.; Díaz Díaz, David
  • Journal of the American Chemical Society, Vol. 129, Issue 42
  • DOI: 10.1021/ja072679d

Benzimidazole and Related Ligands for Cu-Catalyzed Azide−Alkyne Cycloaddition
journal, October 2007

  • Rodionov, Valentin O.; Presolski, Stanislav I.; Gardinier, Sean
  • Journal of the American Chemical Society, Vol. 129, Issue 42
  • DOI: 10.1021/ja072678l

Direct Evidence of a Dinuclear Copper Intermediate in Cu(I)-Catalyzed Azide-Alkyne Cycloadditions
journal, April 2013


On the Mechanism of Copper(I)-Catalyzed Azide-Alkyne Cycloaddition
journal, May 2016

  • Zhu, Lei; Brassard, Christopher J.; Zhang, Xiaoguang
  • The Chemical Record, Vol. 16, Issue 3
  • DOI: 10.1002/tcr.201600002

Experimental Investigation on the Mechanism of Chelation-Assisted, Copper(II) Acetate-Accelerated Azide–Alkyne Cycloaddition
journal, September 2011

  • Kuang, Gui-Chao; Guha, Pampa M.; Brotherton, Wendy S.
  • Journal of the American Chemical Society, Vol. 133, Issue 35
  • DOI: 10.1021/ja203733q

Dicopper Cu(I)Cu(I) and Cu(I)Cu(II) Complexes in Copper-Catalyzed Azide–Alkyne Cycloaddition
journal, April 2017

  • Ziegler, Micah S.; Lakshmi, K. V.; Tilley, T. Don
  • Journal of the American Chemical Society, Vol. 139, Issue 15
  • DOI: 10.1021/jacs.6b13261

Fluorescence Resonance Energy Transfer (FRET) as a High-Throughput Assay for Coupling Reactions. Arylation of Amines as a Case Study
journal, June 2003

  • Stauffer, Shaun R.; Hartwig, John F.
  • Journal of the American Chemical Society, Vol. 125, Issue 23
  • DOI: 10.1021/ja034161p

Optimization of a Biomimetic Transamination Reaction
journal, September 2008

  • Scheck, Rebecca A.; Dedeo, Michel T.; Iavarone, Anthony T.
  • Journal of the American Chemical Society, Vol. 130, Issue 35
  • DOI: 10.1021/ja802495w

Cucurbit[6]uril-Promoted Click Chemistry for Protein Modification
journal, July 2017

  • Finbloom, Joel A.; Han, Kenneth; Slack, Clancy C.
  • Journal of the American Chemical Society, Vol. 139, Issue 28
  • DOI: 10.1021/jacs.7b05164

Discovery of Chemoselective and Biocompatible Reactions Using a High-Throughput Immunoassay Screening
journal, October 2013

  • Kolodych, Sergii; Rasolofonjatovo, Evelia; Chaumontet, Manon
  • Angewandte Chemie International Edition, Vol. 52, Issue 46
  • DOI: 10.1002/anie.201305645

Discovery and Characterization of Catalysts for Azide−Alkyne Cycloaddition by Fluorescence Quenching
journal, August 2004

  • Lewis, Warren G.; Magallon, Fernando G.; Fokin, Valery V.
  • Journal of the American Chemical Society, Vol. 126, Issue 30
  • DOI: 10.1021/ja048425z

Increasing the Efficacy of Bioorthogonal Click Reactions for Bioconjugation: A Comparative Study
journal, July 2011

  • Besanceney-Webler, Christen; Jiang, Hao; Zheng, Tianqing
  • Angewandte Chemie International Edition, Vol. 50, Issue 35
  • DOI: 10.1002/anie.201101817

Biocompatible Copper(I) Catalysts for in Vivo Imaging of Glycans
journal, December 2010

  • Soriano del Amo, David; Wang, Wei; Jiang, Hao
  • Journal of the American Chemical Society, Vol. 132, Issue 47
  • DOI: 10.1021/ja106553e

Quantum Dot Peptide Biosensors for Monitoring Caspase 3 Proteolysis and Calcium Ions
journal, August 2010

  • Prasuhn, Duane E.; Feltz, Anne; Blanco-Canosa, Juan B.
  • ACS Nano, Vol. 4, Issue 9
  • DOI: 10.1021/nn1016132

Labeling Live Cells by Copper-Catalyzed Alkyne−Azide Click Chemistry
journal, October 2010

  • Hong, Vu; Steinmetz, Nicole F.; Manchester, Marianne
  • Bioconjugate Chemistry, Vol. 21, Issue 10
  • DOI: 10.1021/bc100272z

Asymmetric Assembling of Iron Oxide Nanocubes for Improving Magnetic Hyperthermia Performance
journal, November 2017


Rationally Designed Energy Transfer in Upconverting Nanoparticles
journal, March 2015

  • Chan, Emory M.; Levy, Elizabeth S.; Cohen, Bruce E.
  • Advanced Materials, Vol. 27, Issue 38
  • DOI: 10.1002/adma.201500248

Energy-Looping Nanoparticles: Harnessing Excited-State Absorption for Deep-Tissue Imaging
journal, September 2016


Pearson Correlation Analysis of Microarray Data Allows for the Identification of Genetic Targets for Early B-cell Factor
journal, April 2004

  • Månsson, Robert; Tsapogas, Panagiotis; Åkerlund, Mikael
  • Journal of Biological Chemistry, Vol. 279, Issue 17
  • DOI: 10.1074/jbc.M400589200

Correlation of microarray-based breast cancer molecular subtypes and clinical outcomes: implications for treatment optimization
journal, April 2011


Mapping Adverse Drug Reactions in Chemical Space
journal, April 2009

  • Scheiber, Josef; Jenkins, Jeremy L.; Sukuru, Sai Chetan K.
  • Journal of Medicinal Chemistry, Vol. 52, Issue 9
  • DOI: 10.1021/jm801546k

Atomic structure of a voltage-dependent K+ channel in a lipid membrane-like environment
journal, November 2007

  • Long, Stephen B.; Tao, Xiao; Campbell, Ernest B.
  • Nature, Vol. 450, Issue 7168
  • DOI: 10.1038/nature06265

Solution Structure of GxTX-1E, a High-Affinity Tarantula Toxin Interacting with Voltage Sensors in Kv2.1 Potassium Channels,
journal, June 2010

  • Lee, Seungkyu; Milescu, Mirela; Jung, Hyun Ho
  • Biochemistry, Vol. 49, Issue 25
  • DOI: 10.1021/bi100246u

    Works referencing / citing this record:

    Low irradiance multiphoton imaging with alloyed lanthanide nanocrystals
    journal, August 2018

    • Tian, Bining; Fernandez-Bravo, Angel; Najafiaghdam, Hossein
    • Nature Communications, Vol. 9, Issue 1
    • DOI: 10.1038/s41467-018-05577-8

    Simultaneous near-infrared and green fluorescence from single conjugated polymer dots with aggregation-induced emission fluorogen for cell imaging
    journal, January 2018

    • Chen, Sitong; Cui, Shuang; Du, Rongxin
    • Journal of Materials Chemistry B, Vol. 6, Issue 47
    • DOI: 10.1039/c8tb02346h

    The CuAAC: Principles, Homogeneous and Heterogeneous Catalysts, and Novel Developments and Applications
    journal, October 2019

    • Neumann, Steve; Biewend, Michel; Rana, Sravendra
    • Macromolecular Rapid Communications, Vol. 41, Issue 1
    • DOI: 10.1002/marc.201900359

    Ligand-conjugated quantum dots for fast sub-diffraction protein tracking in acute brain slices
    journal, January 2020

    • Thal, Lucas B.; Mann, Victor R.; Sprinzen, David
    • Biomaterials Science, Vol. 8, Issue 3
    • DOI: 10.1039/c9bm01629e

    Electrostatic Repulsion Controls Efficiency of Cu‐Free Click‐Reaction with Azide‐Modified Semiconductor Quantum Dots
    journal, January 2020

    • Ranishenka, Bahdan; Ulashchik, Egor; Radchanka, Aliaksandra
    • ChemNanoMat, Vol. 6, Issue 2
    • DOI: 10.1002/cnma.201900609