Mimetic ligand-based affinity purification of immune complexes and immunoconjugates.
We developed a simple purification method to purify alkaline phosphatase/anti-alkaline phosphatase IgG as immune complexes using mimetic affinity chromatography wherein the antibody was either a monospecific antibody, a bispecific antibody or a commercial polyclonal IgG conjugated with alkaline phosphatase (AP-IgG) covalently. The immune complexes or conjugates were efficiently bound on the mimetic Blue A6XL column and eluted under mild conditions (5-20 mM phosphate buffer). A similar strategy of purifying peroxidase/anti-peroxidase antibody complexes was also successfully demonstrated using the mimetic Red 3 column. Mimetic affinity chromatography thus appears to be a simple method to purify the desired monospecific or bispecific antibodies from the respective hybridomas and quadromas.
Xu
D
D
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada.
Leveugle
B
B
Kreutz
F T
FT
Suresh
M R
MR
D000469:Alkaline Phosphatase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D000936:Antigen-Antibody Complex / Q000276:immunology / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D007106:Immune Sera / Q000276:immunology; D018796:Immunoconjugates / Q000276:immunology / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D008024:Ligands; D051379:Mice; D008807:Mice, Inbred BALB C; D009994:Osmolar Concentration; D051381:Rats; D014407:Tumor Cells, Cultured / Q000276:immunology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007106:Immune Sera; D018796:Immunoconjugates; D007074:Immunoglobulin G; D008024:Ligands; D000469:Alkaline Phosphatase