A label-free and sensitive fluorescence assay for hyaluronidase activity through electrostatic-controlled quantum dots self-assembly

Abstract

A label-free fluorescence assay for hyaluronidase (HAase) activity based on self-assembly of quantum dots is developed. A cationic polymer (polycation) can induce aggregation of the negatively charged quantum dots through electrostatic interactions and the fluorescence of the quantum dots is quenched. When the polycation is mixed with hyaluronic acid (HA), intense binding of HA to the polycation makes the quantum dots free and recovery of the fluorescence of the quantum dots is observed. However, in the presence of HAase, HA is hydrolyzed into small fragments and the polycation induces reaggregation of the quantum dots. A simple and rapid fluorescence sensor with high sensitivity and selectivity for HAase activity detection is therefore successfully established with a detection limit of 0.01 U/mL. Moreover, we have demonstrated an assay that can be applied to detect HAase activity in a complex mixture sample including 1% human serum.