Absence of Protein G-Fc Interaction in Ficin-Derived Mouse IgG1Digests

Typical procedure for IgG fragmentation is based on proteolytic cleavage at the hinge region and usually involves a post-digestion purification step. In mice, IgG1 has been found to bind poorly to protein A. As a result, protein G chromatography could be considered as an alternative for Fc removal. Protein G is generally expected to bind specifically to the Fc region of IgG, but applying protein G for the purification of Fab2 fragment of mouse monoclonal IgG1 under standard physiological conditions, we obtained reproducible clone-independent negligible protein G-Fc reactivity and strong protein G-Fab2 interaction.