title: Stable and Reusable Electrochemical Biosensor for Poly(ADP-ribose) Polymerase and Its Inhibitor Based on Enzyme-Initiated Auto-PARylation.
abstract: A stable and reusable electrochemical biosensor for the label-free detection of poly(ADP-ribose) polymerase (PARP) is designed in this work. C-kit-1, a thiol-modified G-quadruplex oligonucleotide, is first self-assembled on a gold electrode surface. The G-quadruplex structure of c-kit-1 can specifically tether and activate PARP, resulting in the generation of negatively charged poly(ADP-ribose) polymer (PAR). On the basis of electrostatic attraction, PAR facilitates the surface accumulation of positively charged electrochemical signal molecules. Through the characterization of electrochemical signal molecules, the label-free quantification of PARP is simply implemented. On the basis of the proposed method, selective quantification of PARP can be achieved over the linear range from 0.01 to 1 U with a calculated detection limit of 0.003U. Further studies also demonstrate the applicability of the proposed method to biosamples revealing the broad potential in practical applications. Furthermore, inhibitor of PARP has also been detected with this biosensor. Meanwhile, benefited from self-assembly on solid surface, this biosensor possesses two important features, i.e., reusability and stability, which are desirable in related biosensors.
authors.affiliation: Key Laboratory of Animal Physiology and Biochemistry, College of Veterinary Medicine, Nanjing Agricultural University , Nanjing, Jiangsu 210095, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Jiangsu Key Laboratory of Biofunctional Materials, School of Chemistry and Materials Science, Nanjing Normal University , Nanjing, Jiangsu 210023, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory of Biofunctional Materials, School of Chemistry and Materials Science, Nanjing Normal University , Nanjing, Jiangsu 210023, P. R. China.
authors.forename: Zhaoyin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory of Biofunctional Materials, School of Chemistry and Materials Science, Nanjing Normal University , Nanjing, Jiangsu 210023, P. R. China.
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Dai
mesh_terms: D015374:Biosensing Techniques; D000074747:Poly ADP Ribosylation; D011065:Poly(ADP-ribose) Polymerases
chemical_list: D011065:Poly(ADP-ribose) Polymerases
keywords: electrochemical biosensor; enzyme-initiated auto-PARylation; inhibitor; poly(ADP-ribose) polymerase; stability and reusability