An improved method for the purification of IgG monoclonal antibodies from culture supernatants.
A method for the purification of mouse monoclonal antibodies from hybridoma culture supernatants is described. The protocol involves the use of a combination of three previously described methods for the concentration and purification of monoclonal antibodies. Firstly, hybridomas were grown in a Diacult dialysis system (Inter Med Laboratory, Denmark) to yield milligram quantities of monoclonal antibody in a culture supernatant. Monoclonal antibodies were then purified from the culture supernatant by precipitation with polyethylene glycol 6000 (PEG 6000) and finally reprecipitated using an ammonium sulphate procedure. The PEG 6000 treatment caused a density change in the ammonium sulphate immunoglobulin precipitate, and resulted in the formation of a pellicle which contained pure mouse monoclonal antibody. The protocol removed contaminating bovine serum immunoglobulin as well as other serum and cellular protein from the monoclonal antibody preparations.
Brooks
D A
DA
Department of Chemical Pathology, Adelaide Medical Centre for Women and Children, Australia.
Bradford
T M
TM
Hopwood
J J
JJ
D000645:Ammonium Sulfate; D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D011232:Chemical Precipitation; D002803:Chondro-4-Sulfatase / Q000276:immunology; D003470:Culture Media; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008722:Methods; D051379:Mice; D011092:Polyethylene Glycols
D000911:Antibodies, Monoclonal; D003470:Culture Media; D007074:Immunoglobulin G; D011092:Polyethylene Glycols; D002803:Chondro-4-Sulfatase; D000645:Ammonium Sulfate