A label-free conjugated polymer-based fluorescence assay for the determination of adenosine triphosphate and alkaline phosphatase

In this paper, a simple, sensitive, label-free fluorescence sensor for the detection of adenosine triphosphate and alkaline phosphatase was developed, which was based on the water-soluble fluorescent conjugated polymer. Cu2+ could efficiently quench the photoluminescence (PL) intensity of fluorescent conjugated polymer PPESO3 due to the strong electrostatic interaction and electron transfer between PPESO3 and Cu2+. However, the addition of adenosine triphosphate (ATP) could disrupt the polymer–metal complex, leading to the recovery of the fluorescence of PPESO3. The PL intensity ratio I/I0 (I0 and I are the fluorescence intensity of the PPESO3–Cu2+ system in the absence and the presence of ATP, respectively) was proportional to the concentration of ATP. The proposed method was successfully applied to the detection of ATP in human serum samples with satisfactory results. Moreover, considering that ATP could be hydrolyzed by alkaline phosphatase (ALP) and the released Cu2+ could quench the fluorescence of PPESO3, the enzyme activity of ALP was also studied.

[1]  E. Wang,et al.  A carbon nanotubes based ATP apta-sensing platform and its application in cellular assay. , 2010, Biosensors & bioelectronics.

[2]  Cationic conjugated polyelectrolyte-based sensitive fluorescence assay for adenosinetriphosphate and alkaline phosphatase , 2012 .

[3]  C. Yang,et al.  Mass amplifying probe for sensitive fluorescence anisotropy detection of small molecules in complex biological samples. , 2012, Analytical chemistry.

[4]  Guochun Zhao,et al.  Applicability of capillary electrophoresis to the analysis of trace rare earth elements in geological samples. , 2006, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[5]  Synchronous determination of mercury (II) and copper (II) based on quantum dots-multilayer film. , 2011, Analytica chimica acta.

[6]  Yaping Hu,et al.  Graphene signal amplification for sensitive and real-time fluorescence anisotropy detection of small molecules. , 2013, Analytical chemistry.

[7]  Yong-liang Yu,et al.  A new strategy for the detection of adenosine triphosphate by aptamer/quantum dot biosensor based on chemiluminescence resonance energy transfer. , 2012, The Analyst.

[8]  Hong-Wu Tang,et al.  Low background signal platform for the detection of ATP: when a molecular aptamer beacon meets graphene oxide. , 2011, Biosensors & bioelectronics.

[9]  W. Qin,et al.  Potentiometric aptasensing based on target-induced conformational switch of a DNA probe using a polymeric membrane silver ion-selective electrode. , 2013, Biosensors & bioelectronics.

[10]  R. Sakaguchi,et al.  Simultaneous detection of ATP and GTP by covalently linked fluorescent ribonucleopeptide sensors. , 2013, Journal of the American Chemical Society.

[11]  Deqing Zhang,et al.  Continuous on-site label-free ATP fluorometric assay based on aggregation-induced emission of silole. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[12]  B. Mizaikoff,et al.  Local detection of mechanically induced ATP release from bone cells with ATP microbiosensors. , 2013, Biosensors & bioelectronics.

[13]  Mi Hee Kim,et al.  A simple method for improving the optical properties of a dimetallic coordination fluorescent chemosensor for adenosine triphosphate , 2009 .

[14]  M. Jayakannan,et al.  Carboxylic‐functionalized water soluble π‐conjugated polymer: Highly selective and efficient chemosensor for mercury(II) ions , 2009 .

[15]  Wenli Song,et al.  Water-Soluble Iridium(III)-Containing Conjugated Polyelectrolytes with Weakened Energy Transfer Properties for Multicolor Protein Sensing Applications , 2011 .

[16]  Yixiang Cheng,et al.  A highly selective and sensitive polymer-based OFF-ON fluorescent sensor for Hg2+ detection incorporating salen and perylenyl moieties , 2012 .

[17]  F. Feng,et al.  Water-soluble conjugated polymers for continuous and sensitive fluorescence assays for phosphatase and peptidase , 2007 .

[18]  W-C. Lin,et al.  Optical ATP biosensor for extracellular ATP measurement. , 2013, Biosensors & bioelectronics.

[19]  Zhao Li,et al.  A new method for the detection of ATP using a quantum-dot-tagged aptamer , 2008, Analytical and bioanalytical chemistry.

[20]  Hui Yang,et al.  A label-free G-quadruplex-based switch-on fluorescence assay for the selective detection of ATP. , 2012, The Analyst.

[21]  Yun Xiang,et al.  A universal and label-free aptasensor for fluorescent detection of ATP and thrombin based on SYBR Green I dye. , 2013, Biosensors & bioelectronics.

[22]  X. Su,et al.  Optical choline sensor based on a water-soluble fluorescent conjugated polymer and an enzyme-coupled assay , 2013, Microchimica Acta.

[23]  Syed Mazhar Shah,et al.  Water-soluble conjugated polymer–Cu(II) system as a turn-on fluorescence probe for label-free detection of glutathione and cysteine in biological fluids , 2013 .

[24]  Jin Zhu,et al.  Metal ion-sensing polymer in the weak binding monomer regime. , 2009, The journal of physical chemistry. B.

[25]  R. Koncki,et al.  Potentiometric assay for acid and alkaline phosphatase , 2005 .

[26]  F. Liu,et al.  Highly sensitive detection of nitroaromatic explosives using an electrospun nanofibrous sensor based on a novel fluorescent conjugated polymer. , 2012, Analytica chimica acta.

[27]  R. Pereiro,et al.  Fluorescent conjugated polymers for chemical and biochemical sensing , 2011 .

[28]  Yan Liu,et al.  Conjugated polyelectrolyte-based real-time fluorescence assay for alkaline phosphatase with pyrophosphate as substrate. , 2008, Analytical chemistry.

[29]  Jun Wang,et al.  Aptamer-based ATP assay using a luminescent light switching complex. , 2005, Analytical chemistry.

[30]  B. Liu,et al.  Conjugated polyelectrolyte as signal amplifier for fluorogenic probe based enzyme activity study , 2011 .

[31]  Ping Ping Hu,et al.  Carbon nanotubes as a low background signal platform for a molecular aptamer beacon on the basis of long-range resonance energy transfer. , 2010, Analytical chemistry.

[32]  K. Schanze,et al.  A conjugated polyelectrolyte-based fluorescence sensor for pyrophosphate. , 2007, Chemical communications.

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

[34]  Q. Jin,et al.  Fluorescent Conjugated Polymer PPESO3: A Novel Synthetic Route and the Application for Sensing Protease Activities , 2006 .

[35]  A. Kamel,et al.  A simple-potentiometric method for determination of acid and alkaline phosphatase enzymes in biological fluids and dairy products using a nitrophenylphosphate plastic membrane sensor. , 2009, Analytica chimica acta.

[36]  J. Kwak,et al.  Electrochemical determination of total alkaline phosphatase in human blood with a micropatterned ITO film , 2005 .

[37]  Yanchun Zhao,et al.  Mass-amplifying quantum dots in a fluorescence polarization-based aptasensor for ATP , 2013, Microchimica Acta.

[38]  Lidong Li,et al.  Optically amplified DNA detection on self-assembled solid films using conjugated polyelectrolytes , 2012 .

[39]  K. Schanze,et al.  Photophysics, aggregation and amplified quenching of a water-soluble poly(phenylene ethynylene). , 2002, Chemical communications.

[40]  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.

[41]  C. Chung,et al.  Induced self-assembly and Förster resonance energy transfer studies of alkynylplatinum(II) terpyridine complex through interaction with water-soluble poly(phenylene ethynylene sulfonate) and the proof-of-principle demonstration of this two-component ensemble for selective label-free detection of hum , 2011, Journal of the American Chemical Society.

[42]  D. Whitten,et al.  Surface-Enhanced Superquenching of Cyanine Dyes as J-Aggregates on Laponite Clay Nanoparticles , 2002 .