Continuous and sensitive acid phosphatase assay based on a conjugated polyelectrolyte.
We report a novel continuous and sensitive fluorescence turn-on assay for ACPs, which consists of a cationic conjugated polyelectrolyte (PPE4+) and a commonly used phosphatase substrate p-nitrophenyl phosphate (pNPP). The kinetics of the ACP catalyzed hydrolysis of the substrate pNPP was monitored by the fluorescence change of PPE4+ and corresponding kinetic parameters were derived to be consistent with the literature reports. The applications of PPE4+/pNPP-based ACP assay in high-throughput screening of ACP inhibitors and detection of prostatic acid phosphotase (PAP) in vitro were demonstrated.
Yonghua
Y
Xie
Ying
Y
Tan
Renxuan
R
Liu
Rui
R
Zhao
Chunyan
C
Tan
Yuyang
Y
Jiang
D000135:Acid Phosphatase / Q000737:chemistry*; D000814:Aniline Compounds / Q000737:chemistry; D002384:Catalysis; D015195:Drug Design; D004573:Electrolytes / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D008956:Models, Chemical; D009596:Nitrophenols / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry*; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000175:diagnosis; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods
D000814:Aniline Compounds; D004573:Electrolytes; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D011108:Polymers; C008644:nitrophenylphosphate; C068798:4-aminophenylphosphate; D000135:Acid Phosphatase; D010744:Phosphoric Monoester Hydrolases; C428439:prostatic acid phosphatase; D017027:Protein Tyrosine Phosphatases