ABSTRACT
Accomplishment of enzymatic proteolysis via conventional overnight digestion of a therapeutic monoclonal antibody with endoprotease Lys-C was compared to digestion using microwave assisted and pressure cycling technologies.Efficient digestion of proteins using both technologies has been reported in the literature; however there has been limited application to protein therapeutics.

The degree of digestion (number of missed cleavages), and time to achieve complete digestion were investigated for each technique, with conventional digestion used as the control for all experiments performed. Pressure cycling was shown to be most effective at achieving complete digestion in a short time, without unintended perturbation of the molecule. Microwave digestion did not achieve complete digestion and induced oxidation of methionine residues.