Inline ultrafiltration

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.

Section: Introductionmentioning

confidence: 99%

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

Elich¹,

Goodrich²,

Lutz³

et al. 2019

“…While the operation itself is simpler compared to TFF, the sieving behavior is more complicated. In case of completely retained proteins, SPTFF is already in place in industry to provide inline concentration of in‐process pools as reported in literature . One of the major limitations with SPTFF is the need to explore complicated staging arrangements or expand the membrane as higher concentrations are targeted, because the normalized feed flow rates are significantly lower (<10 L/h/m 2 ).…”

Section: Discussionmentioning

confidence: 99%

Ultrafiltration behavior of partially retained proteins and completely retained proteins using equally‐staged single pass tangential flow filtration membranes

Arunkumar

Zhang

Singh

et al. 2018

This work examines the ultrafiltration behavior of partially retained proteins like lysozyme and completely retained proteins like monoclonal antibodies using single pass tangential flow filtration (SPTFF) modules with different screen channels and molecular weight cut-offs. When the staging of the SPTFF used the same membrane area in each stage, there was no impact of the module screened channel or the buffer matrix on the final concentration achieved for completely retained monoclonal antibodies. A hybrid configuration containing 30 kDa membranes and 50 kDa membranes increased the maximum achievable concentration for both the monoclonal antibodies used in this work, at the same time, allowing a two-fold to four-fold increase in normalized feed flow-rate through the system compared to only the 30 kDa or only the 50 kDa membranes. The sieving coefficient of lysozyme measured and calculated using SPTFF was lower than those measured during conventional recirculation TFF indicating a more complicated concentration polarization effect than conventional recirculation TFF. Moreover, the sieving coefficients of lysozyme were the same for the 10 kDa regenerated cellulose and 50 kDa PES membranes while it was higher for the 30 kDa regenerated cellulose membrane. The difference in TFF and SPTFF behavior is important when the product of interest is desired to be permeated. This work presents the first body of data for partially and completely retained solutes together in the SPTFF mode and provides a strategy to increase protein concentration at higher feed flow rates. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018 © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1137-1148, 2018.

“…Recent analysis with a 5 g/L bioreactor titer indicated that buffer/product tank volumes would be the primary bottlenecks in the downstream process . A variety of solutions have been evaluated to address these bottlenecks: inline buffer dilution, in‐line product concentration, high‐capacity Protein A, and no‐salt hydrophobic interaction chromatography . These allow for incremental increases in the capacity of the existing purification trains; however, recent improvements in cell culture processes have allowed companies to achieve these higher titers with a shorter duration in the production bioreactor, forcing downstream development groups to explore ways to purify the product even more efficiently.…”

Section: Introductionmentioning

confidence: 99%

“…Ultrafiltration is utilized at the end of most purification processes to concentrate the product and exchange the product into the desired formulation buffer. It is most commonly operated in batch and fed‐batch mode with the product continuously recirculated; however, recent work has demonstrated that the same membranes can be operated in a single‐pass configuration by connecting multiple filters in series and continuously concentrating the product . We will explore the use of single‐pass tangential flow filtration (SPTFF) to concentrate harvested cell culture fluid (HCCF) and enable frozen storage for process decoupling between upstream and downstream processing.…”

Section: Introductionmentioning

confidence: 99%

Leveraging single‐pass tangential flow filtration to enable decoupling of upstream and downstream monoclonal antibody processing

Brinkmann

Elouafiq

Pieracci

et al. 2018

Decoupling upstream and downstream operations in biopharmaceutical production could enable more flexible manufacturing operations and could allow companies to leverage strategic or financial benefits that would be otherwise unattainable. A decoupling process was developed and scaled up utilizing single-pass tangential flow filtration for volume reduction, followed by bulk freezing in single-use bags prior to purification. Single-pass tangential flow filtration can be used to continuously concentrate harvested cell culture fluid, reducing the volume by 15-25× with a step yield of >96%. These concentration factors were reproduced with a second product, indicating that the process could be amenable to platform processes. Experimental data indicate that the product tested was stable for at least one year at -40 or -70°C. The concentration of the harvested cell culture fluid-either with or without a subsequent period of frozen storage-had no impact on the product quality attributes that were tested. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:405-411, 2018.