title: Overloading ion-exchange membranes as a purification step for monoclonal antibodies.
abstract: The present study examined the overloading of ion-exchange membrane adsorbers, a form of frontal chromatography, as the final purification step in the production of mAbs (monoclonal antibodies) produced from CHO (Chinese-hamster ovary) cells. Preferential binding of impurities over antibody product was exploited using commercially available cation- and anion-exchange membranes. Three different antibody feedstreams previously purified over Protein A and ion-exchange column chromatography were tested. Feedstream conductivity and pH were adjusted to induce product and impurity adsorption. Membranes were then overloaded in a normal flow mode, resulting in retention of impurities and breakthrough of purified antibody. Although some amount of the product also binds to the membranes (usually < or =30 g mAb/l membrane), yields of > or =99% were achieved by marginalizing the losses, typically by loading more than 3 kg mAb/l membrane. Analyses of the purified pools show consistent removal of impurities despite strong mAb-ligand interactions and high membrane loadings. The clearance of host cell proteins was affected by pH and conductivity, but was unaffected by flow rate, membrane properties or scale. The importance of the present study lies in our demonstration of an alternative use of ion-exchange membranes for fast, effective and high yielding purification of mAbs.
authors.affiliation: Process Technical Development, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA. abrown@gene.com
authors.forename: Arick
authors.initials: A
authors.lastname: Brown
authors.forename: Jerome
authors.initials: J
authors.lastname: Bill
authors.forename: Timothy
authors.initials: T
authors.lastname: Tully
authors.forename: Asha
authors.initials: A
authors.lastname: Radhamohan
authors.forename: Chris
authors.initials: C
authors.lastname: Dowd
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004340:Drug Contamination; D004553:Electric Conductivity; D005839:Gentamicins / Q000032:analysis / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007475:Ion Exchange Resins / Q000737:chemistry; D008567:Membranes, Artificial; D013205:Staphylococcal Protein A / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D005839:Gentamicins; D007475:Ion Exchange Resins; D008567:Membranes, Artificial; D013205:Staphylococcal Protein A