The aim of this study was to develop a single-port system that combines glucose monitoring and insulin infusion. In a preclinical proof-of-concept trial, the performance of the glucose measurement at the site of insulin infusion was assessed. Glucose levels were clamped from 40 to 250 mg/dL by intravenous glucose infusion and subcutaneous insulin infusion via the glucose sensor. Sensor-glucose values correlated well with reference blood-glucose values, despite infusion at the site of glucose measurement. The average median ARE value was 21.6 ± 5.7 % for sensors used for insulin infusion, 18.1 ± 5.8 % for sensors used for NaCl infusion and 19.2 ± 7.9 % for sensors without infusion. These preclinical in vivo results demonstrate that single-port glucose monitoring is feasible at the site of insulin infusion.