title: Computer assisted liquid chromatographic method development for the separation of therapeutic proteins.
abstract: This review summarizes the use of computer assisted liquid chromatographic method development for the analytical characterization of protein biopharmaceuticals. Several modes of chromatography including reversed-phase liquid chromatography (RPLC), ion exchange chromatography (IEX), hydrophobic interaction chromatography (HIC) and some perspectives are discussed. For all these chromatographic modes, the most important variables for tuning retention and selectivity are exposed. Then, the retention models that were applied in the literature in RPLC, IEX and HIC are described and critically discussed. Finally, some representative examples of separation of therapeutic proteins and mAbs are shown, to illustrate the possibilities offered by the retention modeling approach. At the end, the reliability of the models was excellent, whatever the chromatographic mode, and the retention time prediction errors were systematically below 2%. In addition, a significant amount of time can be saved during method development and robustness testing.
authors.affiliation: Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Eva
authors.initials: E
authors.lastname: Tyteca
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Veuthey
authors.forename: Gert
authors.initials: G
authors.lastname: Desmet
authors.forename: Davy
authors.initials: D
authors.lastname: Guillarme
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Fekete
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D057927:Hydrophobic and Hydrophilic Interactions; D011506:Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D015203:Reproducibility of Results
chemical_list: D000911:Antibodies, Monoclonal; D011506:Proteins