Absence of protein G-Fc interaction in ficin-derived mouse IgG1 digests

Typical procedure for IgG fragmentation is based on proteolytic cleavage at the hinge region and usually involves a post-digestion purification step. in mice, IgG(1) 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 Fab(2) fragment of mouse monoclonal IgG(1) under standard physiological conditions, we obtained reproducible clone-independent negligible protein G-Fe reactivity and strong protein G-Fab(2) interaction.