Abstract: The aim of this study was to optimize static extraction of pancreatic kallikrein from trypsogen power. Orthogonal test was used for optimization, and the factors included extraction pH value, equilibrium pH value, amount of NaCl during adsorption process, elution pH value, amount of NaCl during elution process and CH3COONa. The optimal conditions for static extraction of pancreatic kallikrein from trypsogen power are as follows： extraction pH value, 6.0; equilibrium pH value, 6.5; 0.02 mol/L NaCl during adsorption process; elution pH value, 6.5; 0.35 mol/L during elution process; and no addition of CH3COONa. The extraction rate was more than 76%. This study provides a theoretical basis for further study on separation and purification of pancreatic kallikrein.