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, 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.
Belenky
Alexander S
AS
Research and Development at Bayer Diagnostics Division, Bayer Diagnostics, Tarrytown, New York 10591, USA. alexander.belenky.b@bayer.com
Wask-Rotter
Elzbieta
E
Sommer
Michael J
MJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001426:Bacterial Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005361:Ficain; D006801:Humans; D015151:Immunoblotting; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification; D007141:Immunoglobulin Fc Fragments / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007202:Indicators and Reagents; D051379:Mice; D020097:Natriuretic Peptide, Brain / Q000276:immunology; D013205:Staphylococcal Protein A; D013972:Thyrotropin / Q000276:immunology
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D013205:Staphylococcal Protein A; D020097:Natriuretic Peptide, Brain; D013972:Thyrotropin; D005361:Ficain