A Novel Enzyme-Responded Controlled Release Electrochemical Biosensor for Hyaluronidase Activity Detection

Abstract

Hyaluronidase (HAase) has been regarded as one of the potential cancer biomarkers. In this study, a novel enzyme-responded controlled release electrochemical biosensor has been constructed for HAase detection. Methylene blue (MB) can be adsorbed on hyaluronic acid (HA) to form macromolecular HA–MB complex first through electrostatic interaction between anionic HA and positively charged MB. The prepared complex cannot be filtered out through centrifugal ultrafiltration, and small electrochemical signal can be recorded from the ultrafiltrate. With the help of HAase, HA–MB complex can be cleaved into small pieces through enzyme digestion. The small pieces of HA–MB complex can be easily released from the macromolecular HA–MB complex to the ultrafiltrate by centrifugal ultrafiltration. Then, the enhanced electrochemical signal from MB can be detected in the above-mentioned ultrafiltrate. The enhanced differential pulse voltammetric response of the system has a linear relationship with HAase concentration in the range of 4.0–60 U·mL −1 , with the limit of detection of 0.28 U·mL −1 . The developed strategy has been successfully applied to detect HAase activity in urine samples.