A pyrophosphatase biosensor with photocurrent analysis

Abstract

A light-induced photochemical biosensor was developed for the analysis of inorganic pyrophosphatase (PPase). PPase hydrolyzes pyrophosphate (PPi) into two independent o-phosphate ions. Two PPi units can chelate a copper ion (Cu2+), forming a PPi-Cu2+-PPi complex and preventing the Cu2+ triggers other reactions. A transparent indium tin oxide (ITO) electrode was coated with a layer of CdS quantum dots (QDs), and then 3,4-diaminobenzoic acid (DBA) was deposited as the anchor. A solution of the PPi-Cu2+-PPi complex and o-phenylenediamine (OPD) was mixed with the analytical sample and then a drop of the mixture was placed on the modified ITO electrode. In the absence of PPase, no reaction occurred between OPD and DBA. A photocurrent was obtained upon excitation of the CdS QDs under light. In the presence of PPase, Cu2+ was released from the complex, triggering the reaction of OPD with DBA on the electrode surface, thereby shielding the CdS QDs from excitation by the light. The observed photocurrent decreased. The difference in the two measured photocurrents corresponded to the activity of PPase. This photochemical biosensor had excellent sensitivity for PPase in the range from 0.8 to 5000 mU, with a limit of detection of 0.41 mU.