Abstract
Peptide CTPR and p‐amino benzamidine (PAB) immobilized on Streamline™ 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.
Acknowledgements
This work was supported by grants from the Fundación Antorchas, the Agencia Nacional de Promoción Científica y Tecnológica de la República Argentina and the Universidad de Buenos Aires.
Silvia A. Camperi, Hector M. Fernandez Lahore, and Osvaldo Cascone are career researchers at the CONICET.