title: Modeling and simulation of anion-exchange membrane chromatography for purification of Sf9 insect cell-derived virus-like particles.
abstract: Recombinant protein-based virus-like particles (VLPs) are steadily gaining in importance as innovative vaccines against cancer and infectious diseases. Multiple VLPs are currently evaluated in clinical phases requiring a straightforward and rational process design. To date, there is no generic platform process available for the purification of VLPs. In order to accelerate and simplify VLP downstream processing, there is a demand for novel development approaches, technologies, and purification tools. Membrane adsorbers have been identified as promising stationary phases for the processing of bionanoparticles due to their large pore sizes. In this work, we present the potential of two strategies for designing VLP processes following the basic tenet of 'quality by design': High-throughput experimentation and process modeling of an anion-exchange membrane capture step. Automated membrane screenings allowed the identification of optimal VLP binding conditions yielding a dynamic binding capacity of 5.7 mg/mL for human B19 parvovirus-like particles derived from Spodoptera frugiperda Sf9 insect cells. A mechanistic approach was implemented for radial ion-exchange membrane chromatography using the lumped-rate model and stoichiometric displacement model for the in silico optimization of a VLP capture step. For the first time, process modeling enabled the in silico design of a selective, robust and scalable process with minimal experimental effort for a complex VLP feedstock. The optimized anion-exchange membrane chromatography process resulted in a protein purity of 81.5%, a DNA clearance of 99.2%, and a VLP recovery of 59%.
authors.affiliation: Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Ladd Effio
authors.affiliation: Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Hahn
authors.affiliation: Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Seiler
authors.affiliation: Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe, Germany; Boehringer Ingelheim Pharma GmbH & Co. KG, Germany.
authors.forename: Stefan A
authors.initials: SA
authors.lastname: Oelmeier
authors.affiliation: Diarect AG, Freiburg, Germany.
authors.forename: Iris
authors.initials: I
authors.lastname: Asen
authors.affiliation: Diarect AG, Freiburg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Silberer
authors.affiliation: Sartorius Stedim Biotech, Göttingen, Germany.
authors.forename: Louis
authors.initials: L
authors.lastname: Villain
authors.affiliation: Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe, Germany. Electronic address: juergen.hubbuch@kit.edu.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hubbuch
mesh_terms: D000818:Animals; D002852:Chromatography, Ion Exchange; D003198:Computer Simulation; D008954:Models, Biological; D011994:Recombinant Proteins / Q000235:genetics; D061987:Sf9 Cells; D058425:Vaccines, Virus-Like Particle / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*
chemical_list: D011994:Recombinant Proteins; D058425:Vaccines, Virus-Like Particle
keywords: Downstream processing; High-throughput process development; Membrane chromatography; Predictive modeling; Quality by design; Virus-like particle vaccine