title: Signal-amplification and real-time fluorescence anisotropy detection of apyrase by carbon nanoparticle.
abstract: Carbon nanomaterial combined with aptamer has been developed as an efficient bioanalytical method in sensor design. Herein, depending on carbon nanoparticle (cCNP)-enhanced fluorescence anisotropy (FA), a novel aptamer-based sensor (aptasensor) enabling signal-amplification and real-time detection of apyrase is reported. The foundation of our sensor design based on ATP-aptamer(P) can be adsorbed on the surface of cCNPs, resulting in the increase of FA due to the mass of cCNPs, and P-ATP complex has weak binding ability to cCNPs with minimal change of FA. Apyrase, being an integral membrane protein, can hydrolyze ATP and make P-ATP complex disassemble, and thus lead to the increasing of FA. Therefore, this approach is demonstrated to be a novel candidate for the detection of apyrase, with high sensitivity and selectivity. The linear dynamic range for the concentrations of apyrase is between 0.1 and 0.5 U/μL along with a detection limit of 0.05 U/μL. Furthermore, these results indicated that our design is a flexible and sensitive method for biomolecule analysis, which makes it promising for practical biomolecule analyses.
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, People's Republic of China; College of Geography and Environmental Science, Zhejiang Normal University, Jinhua 321004, People's Republic of China; Department of Civil Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong, Hong Kong SAR, People's Republic of China. Electronic address: liujh@zjnu.cn.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Liu
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, People's Republic of China; College of Geography and Environmental Science, Zhejiang Normal University, Jinhua 321004, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yu
authors.affiliation: College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, People's Republic of China; College of Geography and Environmental Science, Zhejiang Normal University, Jinhua 321004, People's Republic of China. Electronic address: cjr@zjnu.cn.
authors.forename: Jianrong
authors.initials: J
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Ronghua
authors.initials: R
authors.lastname: Yang
authors.affiliation: Department of Civil Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong, Hong Kong SAR, People's Republic of China. Electronic address: kshih@hku.hk.
authors.forename: Kaimin
authors.initials: K
authors.lastname: Shih
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001081:Apyrase / Q000032:analysis*; D002244:Carbon / Q000737:chemistry*; D003199:Computer Systems; D005454:Fluorescence Polarization / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D012815:Signal Processing, Computer-Assisted; D017550:Spectroscopy, Fourier Transform Infrared; D013997:Time Factors
chemical_list: D002244:Carbon; D000255:Adenosine Triphosphate; D001081:Apyrase
keywords: Aptamer; Apyrase; Carbon nanoparticle; Fluorescence anisotropy; Sensor