title: A fluorometric assay for alkaline phosphatase activity based on β-cyclodextrin-modified carbon quantum dots through host-guest recognition.
abstract: A convenient, reliable and highly sensitive assay for alkaline phosphatase (ALP) activity in the real-time manner is developed based on β-cyclodextrin-modified carbon quantum dots (β-CD-CQDs) nanoprobe through specific host-guest recognition. Carbon quantum dots were first functionalized with 3-aminophenyl boronic acid to produce boronic acid-functionalized CQDs, and then further modified with hydropropyl β-cyclodextrins (β-CD) through B-O bonds to form β-CD-CQDs nanoprobe. p-Nitrophenol phosphate disodium salt is used as the substrate of ALP, and can hydrolyze to p-nitrophenol under the catalysis of ALP. The resulting p-nitrophenol can enter the cavity of β-CD moiety in the nanoprobe due to their specific host-guest recognition, where photoinduced electron transfer process between p-nitrophenol and CQDs takes place to efficiently quench the fluorescence of the probe. The correlation between quenched fluorescence and ALP level can be used to establish quantitative evaluation of ALP activity in a broad range from 3.4 to 100.0U/L with the detection limit of 0.9U/L. This assay shows a high sensitivity to ALP even in the presence of a very high concentration of glucose. This study demonstrates a good electron donor/acceptor pair, which can be used to design general detection strategy through PET process, and also broadens the application of host-guest recognition for enzymes detection in clinical practice.
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Cong
authors.initials: C
authors.lastname: Tang
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Zhaosheng
authors.initials: Z
authors.lastname: Qian
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Jiamin
authors.initials: J
authors.lastname: Xu
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Ao
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Meizhi
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Beijing National laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhou
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China.
authors.forename: Jianrong
authors.initials: J
authors.lastname: Chen
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China. Electronic address: fenghui@zjnu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Feng
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods; D002244:Carbon / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D057075:Enzyme Assays / Q000379:methods; D005470:Fluorometry / Q000379:methods*; D057230:Limit of Detection; D009596:Nitrophenols / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D045663:Quantum Dots / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D047392:beta-Cyclodextrins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D009596:Nitrophenols; D009943:Organophosphorus Compounds; D047392:beta-Cyclodextrins; C008644:nitrophenylphosphate; D002244:Carbon; D000469:Alkaline Phosphatase; C031215:betadex; C024836:4-nitrophenol
keywords: Alkaline phosphatase (ALP) activity; Carbon quantum dots; Photoinduced electron transfer (PET); Real-time fluorometric assay