A working electrode for a subcutaneous sensor for use with a continuous biological monitor for a patient is disclosed. The working electrode includes a conductive substrate and an enzyme layer on the conductive substrate. The enzyme layer includes an enzyme, and the enzyme selected according to a biological function to be monitored. A hydrophobic material cross-linked with an acrylic polyol is included. The enzyme is fully entrapped in the cross-linked hydrophobic material with the acrylic polyol.

The present disclosure provides analyte sensors including one or more NAD(P)-dependent enzymes and an internal supply of NAD(P) for the detection of an analyte. The present disclosure further provides methods of using such analyte sensors for detecting one or more analytes present in a biological sample of a subject, and methods of manufacturing said analyte sensors.

A method for making an enzyme membrane for a working electrode of a continuous biological monitor includes making an aqueous silicone dispersion and making an acrylic polyol emulsion. The silicone dispersion and the acrylic polyol emulsion are mixed to make a base emulsion. An enzyme is added to the base emulsion to create an enzyme/base emulsion dispersion. The enzyme is selected according to a biological function to be monitored. The enzyme/base emulsion dispersion is applied to the working electrode. The applied enzyme/base emulsion dispersion is cured.

The invention provides amperometric analyte sensor systems comprising one or more electrodes designed to monitor in vivo levels of 3-hydroxybutyrate (and optionally glucose as well) in order to facilitate the management of diabetic ketoacidosis. The invention further includes compositions, elements and methods useful with such amperometric analyte sensor systems.

A method for producing 2,4-dihydroxybutyrate (DHB) or L-threonine using a microbial metabolic pathway is disclosed, by expressing the metabolic pathway in a microbial production strain which was previously modified with respect to its natural wild type form by introducing at least one of the genes necessary for the expression of those enzymes used for the enzymatic conversions into the production strain.

The invention provides amperometric analyte sensor systems comprising one or more electrodes designed to monitor in vivo levels of 3-hydroxybutyrate (and optionally glucose as well) in order to facilitate the management of diabetic ketoacidosis. The invention further includes compositions, elements and methods useful with such amperometric analyte sensor systems.