A novel approach to monoclonal antibody separation using high performance liquid affinity chromatography (HPLAC) with SelectiSpher-10 protein G.
Protein G, a bacterial cell wall protein extracted from strains of Streptococci, has been employed as a ligand in high performance liquid affinity chromatography (HPLAC) for separation of monoclonal antibodies. Examples are given of rapid high-resolution separations of rat and mouse monoclonal antibodies belonging to various subclasses. In comparison with protein A chromatography, we were able to show superior binding characteristics for SelectiSpher-10 protein G columns under conditions of 'low' ionic strength (about 0.1 M) and neutral pH (pH approximately 7). The monoclonal antibodies were isolated in high purity (greater than 90%) and with good recovery of specific activity (80-100%). We believe that the HPLAC technology based on SelectiSpher-10 protein G is of potential value in the analysis and purification of monoclonal antibodies from various species and subclasses.
Ohlson
S
S
Perstorp Biolytica AB, Lund, Sweden.
Nilsson
R
R
Niss
U
U
Kjellberg
B M
BM
Freiburghaus
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000378:metabolism; D001666:Binding Sites, Antibody; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007074:Immunoglobulin G / Q000378:metabolism; D007164:Immunosorbents; D007202:Indicators and Reagents; D051379:Mice; D051381:Rats; D013205:Staphylococcal Protein A / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D007074:Immunoglobulin G; D007164:Immunosorbents; D007202:Indicators and Reagents; D013205:Staphylococcal Protein A