Investigating the combination of single‐pass tangential flow filtration and anion exchange chromatography for intensified mAb polishing

Abstract

There is growing interest within the biopharmaceutical industry to improve manufacturing efficiency through process intensification, with the goal of generating more product in less time with smaller equipment. In monoclonal antibody (mAb) purification, a unit operation that can benefit from intensification is anion exchange (AEX) polishing chromatography. Single‐pass tangential flow filtration (SPTFF) technology offers an opportunity for process intensification by reducing intermediate pool volumes and increasing product concentration without recirculation. This study evaluated the performance of an AEX resin, both in terms of host cell protein (HCP) purification and viral clearance, following concentration of a mAb feed using SPTFF. Results show that preconcentration of AEX feed material improved isotherm conditions for HCP binding, resulting in a fourfold increase in resin mAb loading at the target HCP clearance level. Excellent clearance of minute virus of mouse and xenotropic murine virus was maintained at this higher load level. The increased mAb loading enabled by SPTFF preconcentration effectively reduced AEX column volume and buffer requirements, shrinking the overall size of the polishing step. In addition, the suitability of SPTFF for extended processing time operation was demonstrated, indicating that this approach can be implemented for continuous biomanufacturing. The combination of SPTFF concentration and AEX chromatography for an intensified mAb polishing step which improves both manufacturing flexibility and process productivity is supported.

Novel spiking methods developed for anion exchange chromatography operating in a continuous process

“…The viral clearance performance of AEX is influenced by many variables, including the pH and conductivity of the feed and process buffer solutions, load density, flow rate and residence time, all of which have been widely explored in many studies (Miesegaes et al, 2014; Roush, 2014; Strauss et al, 2010; Strauss, Gorrell, et al, 2009; Strauss, Lute, et al, 2009). Eshmuno Q has been shown to achieve consistent levels of viral clearance for MMV (LRV > 5) under the pH (7.5–8.5), conductivity (up to 6 mS/cm), resin load (up to 250 g/L), and residence time (2–8 min) used in this study, and this was confirmed using spiking/loading Method 1 (Elich, Goodrich, Lutz, & Mehta, 2019; Mehta et al, 2016). The current study focused on the impact of potential fluctuations in load and viral spike concentrations that might be introduced by connected, continuous processes, on the viral clearance performance of AEX in flow‐through mode.…”

“…Each bar represents the mean of two runs. MMV, mouse minute virus; TCID50, 50% tissue culture infectious dose [Color figure can be viewed at wileyonlinelibrary.com] to achieve consistent levels of viral clearance for MMV (LRV > 5) under the pH (7.5-8.5), conductivity (up to 6 mS/cm), resin load (up to 250 g/L), and residence time (2-8 min) used in this study, and this was confirmed using spiking/loading Method 1 (Elich, Goodrich, Lutz, & Mehta, 2019;Mehta et al, 2016). The current study focused on the impact of potential fluctuations in load and viral spike concentrations that might be introduced by connected, continuous processes, on the viral clearance performance of AEX in flow-through mode.…”

“…The results also suggest that the batch viral clearance experimental design can potentially be used to evaluate clearance for the AEX flow‐through unit operating in a continuous process. Even though MMV clearance is not sensitive to the pH, resin load and residence time for the mAb feed and Eshmuno Q resin used in this study (Elich et al, 2019; Mehta et al, 2016), future studies covering more extensive operational parameters, including different mAbs, impurity levels, and salt concentrations, are recommended to support this conclusion.…”

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…Apart from these steps, viral inactivation is also used for recombinant biopharmaceuticals produced using mammalian cells (Zydney, 2016). Cell disruption is required for recovery of the desired proteins, expressed as intracellular IBs (Ehgartner et al, 2017). The commonly used cell disruption methods at large scale include high-pressure homogenizers and bead mills (Mevada et al, 2019).…”

Adapting virus filtration to enable intensified and continuous monoclonal antibody processing

“…Single-pass tangential flow filtration can be used to concentrate process intermediates, resulting in potential advantages such as smaller intermediate hold tanks and reduced loading times for subsequent unit operations. 7 These smaller volume, higher concentration intermediates have reduced water and buffer requirements for dilution to achieve compatibility with conditions for downstream purification operations. For polishing chromatography, the mass loading and efficiency of impurity removal is increased due to the higher binding capacity for impurities such as host cell proteins at higher protein concentrations.…”