Separation Of Serine Proteases By para-Aminobenzamidine Affinity Chromatography

Dr Sharon Williams, Product & Downstream Process Development Manager, ProMetic BioSciences Ltd

A diverse variety of chromatographic media can be employed for commercial manufacture of therapeutic biologicals, all of which interact in one way or another with the unique set of characteristics inherent to a protein of interest.

Serine proteases constitute a diverse range of enzymes, defined by the presence of serine residues in their active site and the ability to hydrolyse peptide bonds. They play an important physiological role in blood clotting, inflammation and digestion. Their activity is prone to inhibition by Aminobenzamidine, and immobilisation of this molecule on a solid matrix provides a potent method by which to purify such proteins.

In the manufacture of proteinaceous biopharmaceuticals, capture of proteases can be of benefit because such products are prone to proteolytic degradation and therefore early removal of proteases can improve the overall process yield. 

In this application note the use of immobilised para- Aminobenzamidine is demonstrated for capture and partitioning of two proteases, chymotrypsin and trypsin, on the basis of their differing affinity for the Aminobenzamidine ligand.