Purification of in vitro produced mouse monoclonal antibodies. A two-step procedure utilizing cation exchange chromatography and gel filtration.
A method for production of purified and concentrated mouse monoclonal antibodies was developed. The rationale for the procedure is, firstly, to expand hybridoma cell number in culture medium-containing serum; secondly, to transfer the cells to serum-free medium for production of antibodies; and finally, to harvest antibodies from the conditioned medium by means of cation exchange chromatography on SP-Sephadex C-50, followed by gel filtration on Superose 6B. When compared with chromatography of monoclonal antibodies on protein A-Sepharose our results suggest that the method is particularly useful for purification of antibodies of the IgG1 subclass. Experience from production and purification of 15 various monoclonal antibodies is reported.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000145:classification / Q000302:isolation & purification*; D000918:Antibody Specificity; D000942:Antigens, Bacterial; D002272:Carcinoembryonic Antigen; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006825:Hybridomas; D006863:Hydrogen-Ion Concentration; D007073:Immunoglobulin E; D007526:Isoelectric Point; D051379:Mice; D009344:Neisseria gonorrhoeae / Q000276:immunology; D013972:Thyrotropin
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D002272:Carcinoembryonic Antigen; D007073:Immunoglobulin E; D013972:Thyrotropin