Analysis of interaction between insulin and its receptor by biofunctional chromatography

We used biofunctional chromatography to study the interaction between insulin and immobilized insulin receptor(IR). The heterogeneous insulin receptor extract from Wistar mouse liver was immobilized onto the surface of an immobilized artificial membrane through coating. The detergent was removed by dialysis to reduce its interference with the chromatography. To optimize the protein immobilization, we optimized the coating conditions at: pH 7.2, ion strength 20 mmol, and organic solvent 2% (V/V). The competition and self-displacement interacting study of insulin with IR was conducted by the zonal elution using insulin as the self-competition agent, the result indicated that the insulin receptor was active and interacted with insulin. In addition, relative binding constant was determined by frontal analysis by changing the concentration of insulin in the mobile phase.