Peptide CTPR and p-amino benzamidine (PAB) immobilized on Streamline (TM) were utilized as the chromatographic matrices for trypsin purification from bovine pancreas. By using a clarified pancreas extract, maximum capacity for CTPR-Streamline was 47.4 mg/mL and for PAB-Streamline 78.9 mg/mL while Kd values were 0.39 and 0.38 respectively. Dynamic capacity was 23.0 and 46.0 mg/mL for CTPR- and PAB-Streamline respectively. When the purification process was applied to unclarified pancreas extract in the expanded-bed adsorption mode, 80% trypsin recovery with a purification factor of 18.7 was achieved. Cationic and anionic trypsin obtained from the affinity column were separated by ion-exchange chromatography.