Catalytic Biosensors from Complex Coacervate Core Micelle (C3M) Thin Films.

Enzymes have been applied to a variety of industrially and medically relevant chemistries as both catalysts and sensors. Incorporation of proteins and enzymes into complex coacervates has been demonstrated to improve the thermal, chemical, and temporal stability of enzymes in solution. In this work, a neutral-cationic block copolymer and an enzyme, alkaline phosphatase, are incorporated into complex coacervate core micelles (C3Ms) and coated onto a solid substrate to create a biocatalytic film from aqueous solution. The incorporation of photo-cross-linkable groups into the neutral block of the polymer allows the film to be cross-linked under ultraviolet light, rendering it insoluble. The morphology of the film is shown to depend most strongly on the protein loading within the film, while solvent annealing is shown to have a minimal effect. These films are then demonstrated as specific sensors for Zn2+ in solution in the presence of other metals, a model reaction for ion-selective heavy metal biosensing useful in environmental monitoring. They are shown to have low leaching and maintain sufficient activity and response for sensing for 1 month after aging under ambient conditions and at 40 °C and 50% relative humidity. The C3M immobilization method demonstrated can be applied to a wide variety of proteins with minimal chemical or genetic modification and could be used for immobilization of charged macromolecules in general to produce a wide variety of thin-film devices.