Isolation of Trypsin from Bovine Pancreas Using Immobilized Benzamidine and Peptide CTPR Ligands in Expanded Beds

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.

Keywords:

• Affinity
• expanded bed
• p‐aminobenzamidine
• peptide
• trypsin

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.