title: Isolation of monoclonal antibody from a Chinese hamster ovary supernatant. II: dynamics of the integrated separation on ion exchange and hydrophobic interaction chromatography media.
abstract: Dynamics of the purification process of a CHO derived monoclonal antibody by ion exchange chromatography (IEC), hydrophobic interaction chromatography (HIC) and their integration has been investigated. To quantify the adsorption behavior of the target protein (IgG1) and impurities contained in the supernatant, their elution course on IEC and HIC columns has been analyzed versus pH and/or the salt concentration in the mobile phase. A short-cut method has been proposed for mathematical modeling and determining underlying kinetic and thermodynamic parameters. The accuracy of the model predictions has been verified by comparing the simulated and experimental band profiles recorded in both chromatographic processes. After verification, the model was used to optimize operating conditions for the column loading and chromatographic elution in the integrated process IEC/HIC. Two alternative loading techniques based on the upstream and downstream feed dilution were taken into account in the optimization routine. In the first one the feed stream was diluted with the loading buffer prior to the column loading, while in the latter one the feed dilution was realized inside the column using the multiple-injection technique. It was shown that the downstream dilution allowed significant reduction of the contact time between the protein and the loading buffer.
authors.affiliation: Chemical and Process Engineering Department, Rzeszów University of Technology, Powstańców Warszawy Avenue, 6, 35-959 Rzeszow, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Marek
authors.forename: Renata
authors.initials: R
authors.lastname: Muca
authors.forename: Sylwia
authors.initials: S
authors.lastname: Woś
authors.forename: Wojciech
authors.initials: W
authors.lastname: Piątkowski
authors.forename: Dorota
authors.initials: D
authors.lastname: Antos
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D016466:CHO Cells; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D057927:Hydrophobic and Hydrophilic Interactions; D008962:Models, Theoretical
chemical_list: D000911:Antibodies, Monoclonal
keywords: Hydrophobic interaction chromatography; IgG1; Ion exchange chromatography; Mathematical modeling; Monoclonal antibody