title: Designing new monoclonal antibody purification processes using mixed-mode chromatography sorbents.
abstract: Current platforms for purification of monoclonal antibodies, mostly relying on Protein A as a first capture step, are robust and efficient but significantly increase downstream purification costs, mainly due to Protein A resins. To decrease manufacturing costs, industry is increasingly considering the use of purification schemes without affinity Protein A resins. Mixed-mode chromatography can be used as a powerful alternative to standard purification platforms as it offers new selectivity and separation mechanisms exploiting a combination of both ionic and hydrophobic characteristics of antibodies and contaminating proteins. By using a design of experiments (DoE) approach and high throughput screening in 96-well plates, we developed four different two-steps MAb purification processes, based on the use of mixed-mode sorbents. Finally, three of the tested processes resulted in final purified Mab fractions containing less than 100 ppm of residual CHO proteins (CHOP), with overall process yields above 70%. These data show that mixed-mode chromatography sorbents, used at capture or intermediate purification steps, really expand the options of MAb purification process development with or without Protein A affinity chromatography.
authors: Pall Life Sciences, Cergy St Christophe, France.
authors: Magali
authors: M
authors: Toueille
authors: Audrey
authors: A
authors: Uzel
authors: Jean-François
authors: JF
authors: Depoisier
authors: René
authors: R
authors: Gantier
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D019295:Computational Biology; D006224:Cricetinae; D003412:Cricetulus; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D013205:Staphylococcal Protein A