An Orthogonal Workflow of Electrochemical, Computational, and Thermodynamic Methods Reveals Limitations of Using a Literature-Reported Insulin Binding Peptide in Biosensors.

Developing a continuous insulin-monitoring biosensor is of great importance for both the cellular biomanufacturing industry and for treating diabetes mellitus. Such a sensor needs to be able to effectively monitor insulin across a range of temperatures and pHs and with varying concentrations of competing analytes. One of the two main components of any biosensor is the recognition element, which is responsible for interacting with the molecule of interest. Prior literature describes an insulin-binding peptide (IBP) that was reported to bind to insulin with a 3 nM affinity. Here, we used orthogonal and complementary electrochemical, computational, and thermodynamic characterization methods to evaluate IBP's appropriateness for use in a biosensor. Unfortunately, all three methods failed to produce evidence of IBP-insulin binding either on surfaces or in solution. This indicates that the binding exhibited in previous reports is likely restricted to a limited set of conditions and that IBP is not a suitable recognition element for a continuous insulin biosensor.