Mathematical analysis for impedance change with proteolytic digestion of gelatin.

A mathematical analysis is proposed to demonstrate an inter-relationship between the proteolytic digestion of gelatin on the surface of an interdigitated gold electrode and the resulting rate of impedance change, at different collagenase concentrations, in a biosensor used to detect protease in solution. The impedance change due to digestion of the gelatin layer by collagenase for the overall digestion process was expressed in two different stages: an initial exponential period where the rate of impedance change with enzymic digestion was slow, leading to a critical thickness; after which there was a greater change in impedance associated with subsequent dissolution of the layer and partial or complete uncoating of the digits on the electrode surface. An inter-relationship between the rate of impedance change and collagenase concentration within the range 0.2-0.6 mg ml-1 was predicted for the early stages of the digestion process. A kinetic theory for the rapid rate of impedance change with collagenase concentrations could not be developed owing to the rate remaining almost constant for all concentrations of collagenase, after the critical thickness had been reached. An inter-relationship between the rate of impedance change and stirrer speed was also demonstrated.

Detection of protease activity in the wetted surface of gelatin-coated electrodes in air by AC impedance spectroscopy.

As a step towards developing a biosensor which can detect airborne protease droplets, a biosensor which had previously been developed to detect protease in solution is shown to be capable of detecting different concentrations of protease in liquid films on the sensor surface in air. The biosensor measured impedance change due to proteolytic digestion of its gelatin coating. In saturated air there was a rise in impedance, with a loss in weight of the gelatin, in proportion to collagenase concentration. The addition of glycerol to the gelatin caused a lower impedance response and smaller loss in weight. A critical thickness of the gelatin layer prior to a more rapid change in the rate of impedance was noted, with and without the addition of glycerol. In low air humidity (40%), with gelatin, all collagenase concentrations produced a very similar rapid increase in impedance. However, with glycerol-enhanced gelatin, there was a clear distinction between the extent of impedance change with different collagenase concentrations. The application of these findings for use in the field of bioaerosol sampling is discussed.