Fluorescence‐Amplifying Detection of Hydrogen Peroxide with Cationic Conjugated Polymers, and Its Application to Glucose Sensing

A highly sensitive hydrogen peroxide probe that takes advantage of the amplified fluorescence quenching of conjugated polymers has been developed. The cationic conjugated polymer, poly(9,9-bis(6′-N,N,N-trimethylammonium-hexyl) fluorene phenylene) (PFP-NMe3+) and peroxyfluor-1 with boronate protecting groups (Fl-BB) are used to detect H2O2 optically. Without the addition of H2O2, the absence of electrostatic interactions between the cationic PFP-NMe3+ and the neutral Fl-BB keeps the Fl-BB well separated from the PFP-NMe3+, and no fluorescence quenching of the PFP-NMe3+ occurs. In the presence of H2O2, the formation of the anionic quencher, fluorescein, by specific reaction of the Fl-BB with H2O2 results in strong electrostatic interactions between the PFP-NMe3+ and the fluorescein, and therefore efficient fluorescence quenching of the PFP-NMe3+ occurs. The absorption of fluorescein overlaps the emission of PFP-NMe3+, which encourages fluorescence resonance energy transfer (FRET) from the PFP-NMe3+ to the fluorescein. The H2O2 probe has very good sensitivity, with a detection range of 15 to 600 nM. Since glucose oxidase (GOx) can specifically catalyze the oxidation of β-D-(+)-glucose to generate H2O2, glucose detection is also realized with the H2O2 probe as the signal transducer.

[1]  G. Bazan,et al.  Size‐Specific Interactions Between Single‐ and Double‐Stranded Oligonucleotides and Cationic Water‐Soluble Oligofluorenes , 2003 .

[2]  K. Schanze,et al.  Amplified fluorescence sensing of protease activity with conjugated polyelectrolytes. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Mcbranch,et al.  Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[4]  T. Swager,et al.  Synthesis and application of poly(phenylene ethynylene)s for bioconjugation: a conjugated polymer-based fluorogenic probe for proteases. , 2005, Journal of the American Chemical Society.

[5]  M. Bouchy,et al.  Decolourization of Textile Industry Wastewater by the Photocatalytic Degradation Process , 2001 .

[6]  P. Karplus,et al.  Peroxiredoxin Evolution and the Regulation of Hydrogen Peroxide Signaling , 2003, Science.

[7]  A Heller,et al.  Implanted electrochemical glucose sensors for the management of diabetes. , 1999, Annual review of biomedical engineering.

[8]  G. Bazan,et al.  Collective Optical Behavior of Cationic Water‐Soluble Dendrimers , 2004 .

[9]  R. Haugland,et al.  A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. , 1997, Analytical biochemistry.

[10]  G. Guilbault,et al.  An immunosensor based on the glucose oxidase label and optical oxygen detection. , 1999, Analytical chemistry.

[11]  L Wang,et al.  Raman and FTIR spectroscopies of fluorescein in solutions. , 2001, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[12]  H. Ischiropoulos,et al.  Evaluation of 2',7'-dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells. , 1993, Archives of biochemistry and biophysics.

[13]  Colin L. Masters,et al.  Neurodegenerative diseases and oxidative stress , 2004, Nature Reviews Drug Discovery.

[14]  A. Nègre-Salvayre,et al.  Detection of intracellular reactive oxygen species in cultured cells using fluorescent probes. , 2002, Methods in enzymology.

[15]  G. Bazan,et al.  Optically Amplified RNA–Protein Detection Methods Using Light‐Harvesting Conjugated Polymers , 2003 .

[16]  Daoben Zhu,et al.  Fluorescent amplifying recognition for DNA G-quadruplex folding with a cationic conjugated polymer: a platform for homogeneous potassium detection. , 2005, Journal of the American Chemical Society.

[17]  Tomohiro Sawa,et al.  Chemical basis of inflammation-induced carcinogenesis. , 2003, Archives of biochemistry and biophysics.

[18]  J. Israelachvili.  Intermolecular and surface forces , 1985 .

[19]  P. Worsfold,et al.  Hydrogen peroxide in the marine environment: cycling and methods of analysis , 1992 .

[20]  U. Bunz,et al.  Poly(aryleneethynylene)s: Syntheses, Properties, Structures, and Applications. , 2000, Chemical reviews.

[21]  T. Swager,et al.  Conjugated polymer-based chemical sensors. , 2000, Chemical reviews.

[22]  A. Shah,et al.  Free radicals and redox signalling in cardiovascular disease , 2004, Heart.

[23]  A. Heeger,et al.  Energy Transfer in Mixtures of Water- Soluble Oligomers: Effect of Charge, Aggregation, and Surfactant Complexation , 2002 .

[24]  Ehud Y Isacoff,et al.  A selective, cell-permeable optical probe for hydrogen peroxide in living cells. , 2004, Journal of the American Chemical Society.

[25]  A. Heeger,et al.  Biosensors from conjugated polyelectrolyte complexes , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Guillermo C Bazan,et al.  DNA hybridization detection with water-soluble conjugated polymers and chromophore-labeled single-stranded DNA. , 2003, Journal of the American Chemical Society.

[27]  Qin Zhou,et al.  Fluorescent Chemosensors Based on Energy Migration in Conjugated Polymers: The Molecular Wire Approach to Increased Sensitivity , 1995 .

[28]  T. Swager,et al.  The Molecular Wire Approach to Sensory Signal Amplification , 1998 .

[29]  D. Cremer,et al.  Atmospheric formation of OH radicals and H2O2 from alkene ozonolysis under humid conditions. , 2002, Chemphyschem : a European journal of chemical physics and physical chemistry.

[30]  Alan J. Heeger,et al.  DNA detection using water-soluble conjugated polymers and peptide nucleic acid probes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[31]  W. Rulkens,et al.  Photochemical elimination of phenols and cod in industrial wastewaters , 1997 .

[32]  Michael Schäferling,et al.  Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive europium probe. , 2005, Analytical biochemistry.

[33]  G. Bazan,et al.  Effect of chromophore-charge distance on the energy transfer properties of water-soluble conjugated oligomers. , 2003, Journal of the American Chemical Society.

[34]  Shoko Yoshida,et al.  Fluorescent probes for hydrogen peroxide based on a non-oxidative mechanism. , 2004, Angewandte Chemie.

[35]  G. Bazan,et al.  Fluorescein provides a resonance gate for FRET from conjugated polymers to DNA intercalated dyes. , 2004, Journal of the American Chemical Society.

[36]  F. Wudl,et al.  Highly sensitive biological and chemical sensors based on reversible fluorescence quenching in a conjugated polymer. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[37]  D. Moses,et al.  The fluorescence resonance energy transfer (FRET) gate: a time-resolved study. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Boissinot,et al.  Colorimetric and fluorometric detection of nucleic acids using cationic polythiophene derivatives. , 2002, Angewandte Chemie.

[39]  M. Boissinot,et al.  Fluorescent polymeric transducer for the rapid, simple, and specific detection of nucleic acids at the zeptomole level. , 2004, Journal of the American Chemical Society.

[40]  O. Wolfbeis,et al.  Reversible Optical Sensor Membrane for Hydrogen Peroxide Using an Immobilized Fluorescent Probe, and its Application to a Glucose Biosensor , 2003 .

[41]  Olle Inganäs,et al.  Chip and solution detection of DNA hybridization using a luminescent zwitterionic polythiophene derivative , 2003, Nature materials.

[42]  Qingbo Xu,et al.  Activation of Mitogen-activated Protein Kinase by HO , 1996, The Journal of Biological Chemistry.

[43]  T. Swager,et al.  Signal Amplification of a “Turn-On” Sensor: Harvesting the Light Captured by a Conjugated Polymer , 2000 .

[44]  S. Daunert,et al.  Fluorescence Glucose Detection: Advances Toward the Ideal In Vivo Biosensor , 2004, Journal of Fluorescence.

[45]  Zhihong Lin,et al.  A europium-ion-based luminescent sensing probe for hydrogen peroxide. , 2002, Angewandte Chemie.