Ion exchange resins for the purification of monoclonal antibodies from animal cell culture.
We have compared various ion exchangers for monoclonal antibody (MAb) purification using different starting materials such as ascitic fluid and cell culture supernatant. Twelve cation and anion exchange resins were tested so far. Purification of MAbs with regard to the starting material is described. In well-defined conditions of adsorption (20 mM MES buffer, pH 6.50), one purification step based on cation-exchange chromatography is generally sufficient to achieve at least 90% purity of the MAb, even when produced by animal cell culture. Cation-exchange supports exhibit higher capacity for MAbs compared to anion exchangers. Among the cation exchangers tested, we have selected the cross-linked matrix S Sepharose FF for its large specificity and capacity for MAbs. Considering these key parameters and also the good mechanical resistance of the S Sepharose FF, we describe how, by varying the flow rate, sample concentration, and size of the column, the productivity may be improved in a monoclonal antibody purification process. Finally, a general 'gram scale' purification protocol of MAbs produced by animal cell cultures is proposed. This protocol, based on economical adsorption conditions and three steps of elution (100 mM, 200 mM and 1 M NaCl), allows the recovery of highly purified MAbs.
Graf
H
H
Setrie Génie Industriel, S.G.I., Toulouse, France.
Rabaud
J N
JN
Egly
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001709:Biotechnology; D002021:Buffers; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange / Q000379:methods; D003470:Culture Media; D005069:Evaluation Studies as Topic; D006825:Hybridomas / Q000276:immunology; D006863:Hydrogen-Ion Concentration; D007475:Ion Exchange Resins; D051379:Mice; D016853:Muromonab-CD3 / Q000302:isolation & purification; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000276:immunology
D000911:Antibodies, Monoclonal; D002021:Buffers; D003470:Culture Media; D007475:Ion Exchange Resins; D016853:Muromonab-CD3; D017412:Ribonucleoprotein, U1 Small Nuclear