An alternative downstream process based on aqueous two-phase extraction for the purification of monoclonal antibodies

Kruse, Thomas; Kampmann, Markus; Rüddel, I.; Greller, Gerhard

doi:10.1016/j.bej.2020.107703

Cited by 24 publications

(10 citation statements)

References 54 publications

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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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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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“…Indeed, many traditional polymer-based ABSs have been engineered to successfully purify many APIs, including monoclonal antibodies. [190][191][192][193] Because some ILs and DESs are bioderived, renewable, and their syntheses more economical than those of polymers used in ABSs, [194] it is rational to assume that an IL-or DES-based ABS would be a more sustainable option, especially at large scale. [188,195] Also, using ILs and DESs as phase-forming components in ABSs opens new opportunities to tune the phase's properties, including magnetism, chirality, polarity, and pH, to selectively extract and purify specialty molecules from their native environment.…”

Section: Methodsmentioning

confidence: 99%

Enabling Sustainable Chemistry with Ionic Liquids and Deep Eutectic Solvents: A Fad or the Future?

et al. 2022

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Ionic liquids (ILs) and deep eutectic solvents (DESs) debuted with a promise of a superior sustainability footprint due to their low vapor pressure. However, their toxicity and high cost compromise this footprint, impeding their real‐world applications. Fortunately, their property tunability through a rational selection of precursors, including bioderived ones, provides a strategy to ameliorate toxicity, lower cost, and endow new functions. This Review discusses whether ILs and DESs are sustainable solvents and how they contribute to sustainable chemical processes.

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“…The mAb concentration was determined by an analytical high performance liquid chromatography (HPLC) system with an SEC column [ 34 ].…”

Section: Methodsmentioning

confidence: 99%

A novel approach for enumeration of extracellular vesicles from crude and purified cell culture samples

Kruse

Schneider

Reger

et al. 2022

Engineering in Life Sciences

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The interest in extracellular vesicles (EVs) has been increased in recent years due to their potential application in diagnosis and therapy of severe diseases. The versatile fields of application due to the numerous possible cargos and the targeted delivery system make them a promising biopharmaceutical product. However, their broad size range as well as varied surface protein content result in challenges for the purification, characterization, and quantification. In this study a novel method, based on high-resolution flow cytometry, was examined for the enumeration of EVs in purified as well as crude process samples. In addition to quantification, samples were characterized by dynamic light scattering, zeta potential measurement, and analytical size exclusion chromatography. It has been demonstrated that EVs were successfully enumerated with the novel method, offering great benefits for development and monitoring of EV processes.

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An alternative downstream process based on aqueous two-phase extraction for the purification of monoclonal antibodies

Cited by 24 publications

References 54 publications

Rapid intensification of an established CHO cell fed‐batch process

Rapid intensification of an established CHO cell fed‐batch process

Enabling Sustainable Chemistry with Ionic Liquids and Deep Eutectic Solvents: A Fad or the Future?

A novel approach for enumeration of extracellular vesicles from crude and purified cell culture samples

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