Towards the Development of an Insulin Degradation Test.

People with diabetes rely on exogenous insulin to reduce blood glucose levels, compensating for insulin resistance or impaired pancreatic β-cell function. Despite being essential for diabetes management, insulin formulations exhibit inconsistent performance due to their relatively fragile stability. This instability carries significant cost implications: Some individuals spend over $1000 per month on insulin, and these high prices influence one in six Americans with diabetes to ration their insulin supplies. Environmental stressors can induce conformational changes that cause insulin to misfold and aggregate into fibrils, which are inactive structures that contribute to long-term diabetic complications. Although insulin's instability is well-documented, no test currently exists outside of laboratory settings to determine whether an insulin formulation has fibrillated. Here, we compare biochemical techniques for assessing bioactivity and structural integrity in three commercial insulin analogs exposed to physiologically relevant stress conditions, showing that fibril formation precedes measurable loss of bioactivity in insulin and that fibrillation depends on both the stressor type and the insulin formulation tested. We then demonstrate proof-of-concept testing for antibody-based fibrillation detection using commercial monoclonal antibody candidates. Together, these findings underscore the critical need for accessible insulin quality testing and demonstrate the feasibility of antibody-based detection of insulin fibrillation.