High specificity and efficiency electrochemical detection of poly(ADP-ribose) polymerase-1 activity based on...

Poly(ADP-ribose) polymerase-1 (PARP-1) activity is closely related to tumor, which is a promising biomarker for cancer diagnosis. So far, only a few methods have been developed for PARP-1 activity assay because both PARP-1 and its catalytic products lack valuable optical or electrochemical property. Herein, we propose a more specific method to label probes on great deal of phosphate groups of PAR. Firstly, versatile peptides were used to prepare CuNPs. This peptide not only worked as reducing agent to prepare CuNPs but also had guanidine groups to label PAR autonomously and specifically. Unlike most previously reported methods based on unspecific electrostatic interactions, CuNPs probes covered by guanidine groups labelled PAR with phosphate groups via intense covalent-like interactions. On the other hand, PARP-1 catalyzed the formation of PAR in each isolated reaction container of the detection array, realizing the high-throughput detection and enhancing the detection efficiency. Ultimately, CuNPs were oxidized into Cu²⁺ and precisely detected by stripping voltammetry. Hence, selectivity and efficiency of PARP-1 detection were both improved. Meanwhile, this approach was successfully used to detect the efficiency of PARP-1 inhibitor and the PARP-1 contents in real cells, indicating its great potential for clinical diagnosis and high-throughput PARP-1 inhibitor screen.