title: Maximizing binding capacity for protein A chromatography.
abstract: Advances in cell culture expression levels in the last two decades have resulted in monoclonal antibody titers of ≥10 g/L to be purified downstream. A high capacity capture step is crucial to prevent purification from being the bottleneck in the manufacturing process. Despite its high cost and other disadvantages, Protein A chromatography still remains the optimal choice for antibody capture due to the excellent selectivity provided by this step. A dual flow loading strategy was used in conjunction with a new generation high capacity Protein A resin to maximize binding capacity without significantly increasing processing time. Optimum conditions were established using a simple empirical Design of Experiment (DOE) based model and verified with a wide panel of antibodies. Dynamic binding capacities of >65 g/L could be achieved under these new conditions, significantly higher by more than one and half times the values that have been typically achieved with Protein A in the past. Furthermore, comparable process performance and product quality was demonstrated for the Protein A step at the increased loading.
authors.affiliation: Dept. of Process Biochemistry, Bioprocess Development, Biogen Idec, 5000 Davis Drive, Research Triangle Park, NC.
authors.forename: Sanchayita
authors.initials: S
authors.lastname: Ghose
authors.forename: Jennifer
authors.initials: J
authors.lastname: Zhang
authors.forename: Lynn
authors.initials: L
authors.lastname: Conley
authors.forename: Ryan
authors.initials: R
authors.lastname: Caple
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Williams
authors.forename: Douglas
authors.initials: D
authors.lastname: Cecchini
mesh_terms: D000465:Algorithms; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000378:metabolism*; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D011485:Protein Binding; D012107:Research Design; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D013205:Staphylococcal Protein A