Fluorescence Regulation of Poly(thymine)-Templated Copper Nanoparticles via an Enzyme-Triggered Reaction toward Sensitive and Selective Detection of Alkaline Phosphatase.

The activity of alkaline phosphatase (ALP) is a crucial index of blood routine examinations, since the concentration of ALP is highly associated with various human diseases. To address the demands of clinical tests, efforts should be made to develop more approaches that can sense ALP in real samples. Recently, we find that fluorescence of poly(30T)-templated copper nanoparticles (CuNPs) can be directly and effectively quenched by pyrophosphate ion (PPi), providing new perspective in designing sensitive biosensors based on DNA-templated CuNPs. In addition, it has been confirmed that phosphate ion (Pi), product of PPi hydrolysis, does not affect the intense fluorescence of CuNPs. Since ALP can specifically hydrolyze PPi into Pi, fluorescence of CuNPs is thus regulated by an ALP-triggered reaction, and a novel ALP biosensor is successfully developed. As a result, ALP is sensitively and selectively quantified with a wide linear range of 6.0 × 10-2 U/L to 6.0 × 102 U/L and a low detection limit of 3.5 × 10-2 U/L. Besides, two typical inhibitors of ALP are evaluated by this analytical method, and different inhibitory effects are indicated. More importantly, by challenging this biosensor with real human serums, the obtained results get a fine match with the data from clinical tests, and the serum sample from a patient with liver disease is clearly distinguished, suggesting promising applications of this biosensor in clinical diagnosis.

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

[2]  Hai-Bo Wang,et al.  A label-free and ultrasensitive fluorescent sensor for dopamine detection based on double-stranded DNA templated copper nanoparticles , 2015 .

[3]  Yongning Wu,et al.  Facile and Sensitive Fluorescence Sensing of Alkaline Phosphatase Activity with Photoluminescent Carbon Dots Based on Inner Filter Effect. , 2016, Analytical chemistry.

[4]  T. Bigioni,et al.  Glutathione-stabilized magic-number silver cluster compounds. , 2010, Journal of the American Chemical Society.

[5]  L. Thompson,et al.  Therapeutic application of histone deacetylase inhibitors for central nervous system disorders , 2008, Nature Reviews Drug Discovery.

[6]  Linling Zhu,et al.  Triplex molecular beacons for sensitive recognition of melamine based on abasic-site-containing DNA and fluorescent silver nanoclusters. , 2015, Chemical communications.

[7]  Xuemei Wang,et al.  Alkaline phosphatase-responsive anodic electrochemiluminescence of CdSe nanoparticles. , 2012, Analytical chemistry.

[8]  E. Wang,et al.  Photoinduced electron transfer of DNA/Ag nanoclusters modulated by G-quadruplex/hemin complex for the construction of versatile biosensors. , 2013, Journal of the American Chemical Society.

[9]  W. Tan,et al.  DNA-templated in situ growth of silver nanoparticles on mesoporous silica nanospheres for smart intracellular GSH-controlled release. , 2015, Chemical communications.

[10]  Wenying Li,et al.  Nucleic acid-regulated perylene probe-induced gold nanoparticle aggregation: a new strategy for colorimetric sensing of alkaline phosphatase activity and inhibitor screening. , 2014, ACS applied materials & interfaces.

[11]  K. Artyushkova,et al.  A Hybrid DNA-Templated Gold Nanocluster For Enhanced Enzymatic Reduction of Oxygen. , 2015, Journal of the American Chemical Society.

[12]  Shaojun Dong,et al.  Label-free and enzyme-free platform for the construction of advanced DNA logic devices based on the assembly of graphene oxide and DNA-templated AgNCs. , 2016, Nanoscale.

[13]  A. Woolley,et al.  DNA-templated nickel nanostructures and protein assemblies. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[14]  E. Wang,et al.  A nanocluster beacon based on the template transformation of DNA-templated silver nanoclusters. , 2016, Chemical communications.

[15]  G. Liang,et al.  Enzymatic Hydrogelation-Induced Fluorescence Turn-Off for Sensing Alkaline Phosphatase in Vitro and in Living Cells. , 2015, Analytical chemistry.

[16]  Ji Hoon Lee,et al.  Rapid monitoring of alkaline phosphatase in raw milk using chemiluminescence detection. , 2011, Analytical methods : advancing methods and applications.

[17]  Kemin Wang,et al.  Concatemeric dsDNA-templated copper nanoparticles strategy with improved sensitivity and stability based on rolling circle replication and its application in microRNA detection. , 2014, Analytical chemistry.

[18]  Jun Li,et al.  Ratiometric fluorescent probe for alkaline phosphatase based on betaine-modified polyethylenimine via excimer/monomer conversion. , 2014, Analytical chemistry.

[19]  Nick Bishop,et al.  Enzyme-replacement therapy in life-threatening hypophosphatasia. , 2012, The New England journal of medicine.

[20]  Chuan-Hua Zhou,et al.  Reliable Digital Single Molecule Electrochemistry for Ultrasensitive Alkaline Phosphatase Detection. , 2016, Analytical chemistry.

[21]  Zhimin Li,et al.  Protein-directed solution-phase green synthesis of BSA-conjugated M(x)Se(y) (M = Ag, Cd, Pb, Cu) nanomaterials. , 2011, Chemistry, an Asian journal.

[22]  Andriy Mokhir,et al.  Selective dsDNA-templated formation of copper nanoparticles in solution. , 2010, Angewandte Chemie.

[23]  Yongming Guo,et al.  Fluorescent copper nanoparticles: recent advances in synthesis and applications for sensing metal ions. , 2016, Nanoscale.

[24]  Kemin Wang,et al.  Poly(thymine)-templated selective formation of fluorescent copper nanoparticles. , 2013, Angewandte Chemie.

[25]  Manuel Miró,et al.  High-resolution colorimetric assay for rapid visual readout of phosphatase activity based on gold/silver core/shell nanorod. , 2014, ACS applied materials & interfaces.

[26]  Akhtar Hayat,et al.  Nanoceria particles as catalytic amplifiers for alkaline phosphatase assays. , 2013, Analytical chemistry.

[27]  Xin Wu,et al.  Copper-Mediated DNA-Scaffolded Silver Nanocluster On-Off Switch for Detection of Pyrophosphate and Alkaline Phosphatase. , 2016, Analytical chemistry.

[28]  Wei Wang,et al.  Detection of alkaline phosphatase using surface-enhanced Raman spectroscopy. , 2006, Analytical chemistry.

[29]  S. D. Gilman,et al.  Studies of reversible inhibition, irreversible inhibition, and activation of alkaline phosphatase by capillary electrophoresis. , 2002, Analytical biochemistry.

[30]  M. Whyte.  Hypophosphatasia — aetiology, nosology, pathogenesis, diagnosis and treatment , 2016, Nature Reviews Endocrinology.

[31]  K. Shiraki,et al.  High‐molecular intestinal alkaline phosphatase in chronic liver diseases , 2007, Journal of clinical laboratory analysis.

[32]  Xue Zhu,et al.  Rational design of signal-on biosensors by using photoinduced electron transfer between Ag nanoclusters and split G-quadruplex halves-hemin complexes. , 2014, Chemical communications.

[33]  Jianhui Jiang,et al.  Inhibition of dsDNA-templated copper nanoparticles by pyrophosphate as a label-free fluorescent strategy for alkaline phosphatase assay. , 2013, Analytical chemistry.

[34]  Lingwen Zeng,et al.  A simple and sensitive sensor for rapid detection of sulfide anions using DNA-templated copper nanoparticles as fluorescent probes. , 2012, The Analyst.