A novel clarification approach for intensified monoclonal antibody processes with 100 million cells/mL using a single-use fluidized bed centrifuge

Saballus, Martin; Nisser, Lucy; Kampmann, Markus; Greller, Gerhard

doi:10.1016/j.bej.2020.107887

Cited by 17 publications

(32 citation statements)

References 14 publications

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“…The slight increase in turbidity and the associated decrease in CE may be due to the shorter residence time of the cell suspension and therefore a reduction in sedimented particles [7]. Nevertheless, the CE values of 96.6-97.5 % achieved are very high and in agreement with other cell separation methods described in the literature [6,14,[19][20][21]. Minow et al [6] achieved a turbiditybased CE of 96.4-98.0 % when clarifying a CHO cell suspension with depth filters with or without diatomaceous body-feed filters.…”

Section: Resultssupporting

confidence: 85%

“…Through direct washing, a 100 % mAb recovery would theoretically be possible with the kSep. However, Saballus et al [20] showed that only 93.2-97.0 % of the mAb was obtained when using the kSep including an integrated washing process. The mAb concentration decreased due to washing and, thus, a lot of buffer would be necessary for complete mAb recovery.…”

Section: Resultsmentioning

confidence: 99%

“…Minow et al [6] achieved a turbiditybased CE of 96.4-98.0 % when clarifying a CHO cell suspension with depth filters with or without diatomaceous body-feed filters. When separated with the single-use fluidized bed centrifuge kSep, CE values of 90.2-93.4 and 97.5-98.1 %, respectively, were described [14,20]. Regardless of the separation parameters used and the slightly decreasing turbidity-based CE depending on the separation setting, no cells or cell debris in the form of a solid pellet could be detected in the centrate, even after a further discontinuous centrifugation step.…”

Section: Resultsmentioning

confidence: 99%

“…While mechanical seals have a limited lifetime, the completely bearingless drive technology in the kytero 500 allows the rotating parts to last for many days, depending on the bowl speed [17]. Both companies offer their single-use disk stack centrifuges in two versions; the kytero 500 and the CultureOne Primo are recommended for cell harvesting of bioreactor sizes up to 500 L and are thus used on a pilot-scale [19,20]. The kytero 2000, which was launched in October 2022, and the CultureOne Maxi are suitable for production scale and can process cell suspension volumes of up to 2000 L [16].…”

Section: Introductionmentioning

confidence: 99%

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Qualification of a Single‐Use Disk Stack Separator for Cell Separation in Mammalian Cell‐Based Antibody Production

Ott

Müller

Schirmer

et al. 2022

Chemie Ingenieur Technik

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Single‐use technology is now an indispensable part of biopharmaceutical production processes. This not only applies to research and development, but also to commercial processes. The manufacturers of single‐use systems are working on improving them for use in intensified processes as well as on new developments. One of the bottlenecks was the availability of single‐use disk stack separators whose stainless steel variants are widely used in industry. In this study, the suitability of the new single‐use disk stack separator kytero 500 was investigated for the first time for cell separation in pilot‐scale Chinese hamster ovary cell‐based antibody production. The cells were completely separated over a wide range of separator settings with nearly no release of intracellular substances and no loss of the antibody.

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Section: Resultssupporting

confidence: 85%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Qualification of a Single‐Use Disk Stack Separator for Cell Separation in Mammalian Cell‐Based Antibody Production

Ott

Müller

Schirmer

et al. 2022

Chemie Ingenieur Technik

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“…Final samples of the raw supernatant of the Ambr®250 experiments were also subjected for analysis of host cell impurities with respect to host cell proteins (HCP) using an HCP‐ELISA kit (CYG‐F550, Cygnus Technologies) 26 and extracellular DNA assay using the Quant‐iT™ PicoGreen™ dsDNA Assay Kit (ThermoFisher Scientific, Waltham). 27 …”

Section: Methodsmentioning

confidence: 99%

Rapid intensification of an established CHO cell fed‐batch process

Schulze

Niemann

Wijffels

et al. 2021

Biotechnology Progress

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Currently, the mammalian biomanufacturing industry explores process intensification (PI) to meet upcoming demands of biotherapeutics while keeping production flexible but, more importantly, as economic as possible. However, intensified processes often require more development time compared with conventional fed-batches (FBs) preventing their implementation. Hence, rapid and efficient, yet straightforward strategies for PI are needed. In this study we demonstrate such a strategy for the intensification of an N-stage FB by implementing N-1 perfusion cell culture and high inoculum cell densities resulting in a robust intensified FB (iFB). Furthermore, we show successful combination of such an iFB with the addition of productivity enhancers, which has not been reported so far. The conventional CHO cell FB process was step-wise improved and intensified rapidly in multi-parallel small-scale bioreactors using N-1 perfusion. The iFBs were performed in 15 and 250 ml bioreactors and allowed to evaluate the impact on key process indicators (KPI): the space-time yield (STY) was successfully doubled from 0.28 to 0.55 g/L d, while product quality was maintained. This

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Cost‐efficient cell clarification using an intensified fluidized bed centrifugation platform approach

Saballus,

Filz,

Pollard

et al. 2023

Biotech & Bioengineering

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The intensification of industrial biopharmaceutical production and the integration of process steps pave the way for patients to access affordable treatments. The predominantly batchwise biomanufacturing of established cell clarification technologies, stainless steel disc stack centrifugation (DSC) and single‐use (SU) depth filtration (DF), pose technological and economical bottlenecks, that include low biomass loading capacities and low product recoveries. Therefore, a novel SU‐based clarification platform was developed combining fluidized bed centrifugation (FBC) with integrated filtration. The feasibility of this approach was investigated for high cell concentration with more than 100E6 cells/mL. Furthermore, scalability to 200 L bioreactor scale was tested for moderate cell concentrations. In both trials, low harvest turbidities (4 NTU) and superior antibody recoveries (95%) were achieved. The impact on the overall economics of industrial SU biomanufacturing using an up‐scaled FBC approach was compared with DSC and DF technologies for different process parameters. As a result, the FBC showed to be the most cost‐effective alternative for annual mAb production below 500 kg. In addition, the FBC clarification of increasing cell concentrations was found to have minimal impact on overall process costs, in contrast to established technologies, demonstrating that the FBC approach is particularly suitable for intensified processes.

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A novel clarification approach for intensified monoclonal antibody processes with 100 million cells/mL using a single-use fluidized bed centrifuge

Cited by 17 publications

References 14 publications

Qualification of a Single‐Use Disk Stack Separator for Cell Separation in Mammalian Cell‐Based Antibody Production

Qualification of a Single‐Use Disk Stack Separator for Cell Separation in Mammalian Cell‐Based Antibody Production

Rapid intensification of an established CHO cell fed‐batch process

Cost‐efficient cell clarification using an intensified fluidized bed centrifugation platform approach

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