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  1. Article ; Online: Platform development for high-throughput optimization of perfusion processes: Part I: Implementation of cell bleeds in microwell plates.

    Dorn, Marie / Klottrup-Rees, Kerensa / Lee, Ken / Micheletti, Martina

    Biotechnology and bioengineering

    2024  

    Abstract: The promise of continuous processing to increase yields and improve product quality of biopharmaceuticals while decreasing the manufacturing footprint is transformative. Developing and optimizing perfusion operations requires screening various parameters, ...

    Abstract The promise of continuous processing to increase yields and improve product quality of biopharmaceuticals while decreasing the manufacturing footprint is transformative. Developing and optimizing perfusion operations requires screening various parameters, which is expensive and time-consuming when using benchtop bioreactors. Scale-down models (SDMs) are the most feasible option for high-throughput data generation and condition screening. However, new SDMs mimicking perfusion are required, enabling experiments to be run in parallel. In this study, a method using microwell plates (MWP) operating in semi-perfusion mode with an implemented cell bleed step is presented. A CHO cell line was cultivated in a 24-well MWP (V
    Language English
    Publishing date 2024-02-23
    Publishing country United States
    Document type Journal Article
    ZDB-ID 280318-5
    ISSN 1097-0290 ; 0006-3592
    ISSN (online) 1097-0290
    ISSN 0006-3592
    DOI 10.1002/bit.28682
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.B 960: Show issues Location:
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  2. Article ; Online: Development of mL-scale pseudo-perfusion methodologies for high-throughput early phase development studies

    Tregidgo, Molly / Lucas, Ciara / Dorn, Marie / Micheletti, Martina

    Biochemical Engineering Journal. 2023 June, v. 195 p.108906-

    2023  

    Abstract: Small-scale devices enable rapid development and optimization, and while many exist for fed-batch cultures, there are few comparable devices for high cell density perfusion cultures. To address this gap, pseudo-perfusion methodologies were developed in ... ...

    Abstract Small-scale devices enable rapid development and optimization, and while many exist for fed-batch cultures, there are few comparable devices for high cell density perfusion cultures. To address this gap, pseudo-perfusion methodologies were developed in microwell plates at mL-scale capable of achieving many of the specific characteristics of perfusion culture, including high cell density, cell retention and increased productivity. Pseudo-perfusion was achieved via sedimentation or centrifugation of 24 microwell plates prior to media exchange 1-2 times daily, generating separation efficiencies higher than 90%. Media exchanges commenced on day 3 and achieved perfusion rates of 0.5-1.8 vessel volumes per day (VVD). Pseudo-perfusion methodologies resulted in maximum viable cell densities (VCDs) of up to 42 × 10⁶ cells mL⁻¹, 4.2-fold greater than fed-batch cultures. Volumetric productivities increased by 1.9-fold, generating industrially relevant productivities. Similar performance was observed between sedimentation and centrifugation methodologies, with minor deviations attributed to elongated manipulation times and lower packed cell density prior to separation when sedimentation was used. The microwell plate (MWP) experiments were validated at the 5 L scale and showed comparability in metabolite and growth profiles. The work presents the development of a robust tool for high-throughput development studies which is sensitive to changes in media composition and exchange rate, which could be reliably used for initial screening of high cell density perfusion cultures.
    Keywords cell viability ; centrifugation ; metabolites ; CHO, ; CSPR ; mAbs, Viable cell density ; VVD ; CHO cells ; Microscale ; Perfusion culture ; Monoclonal antibodies ; High throughput
    Language English
    Dates of publication 2023-06
    Publishing place Elsevier B.V.
    Document type Article ; Online
    Note Use and reproduction
    ZDB-ID 2012139-8
    ISSN 1369-703X
    ISSN 1369-703X
    DOI 10.1016/j.bej.2023.108906
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Platform development for high-throughput optimization of perfusion processes-Part II: Variation of perfusion rate strategies in microwell plates.

    Dorn, Marie / Lucas, Ciara / Klottrup-Rees, Kerensa / Lee, Ken / Micheletti, Martina

    Biotechnology and bioengineering

    2024  

    Abstract: The biopharmaceutical industry is replacing fed-batch with perfusion processes to take advantage of reduced capital and operational costs due to the operation at high cell densities (HCD) and improved productivities. HCDs are achieved by cell retention ... ...

    Abstract The biopharmaceutical industry is replacing fed-batch with perfusion processes to take advantage of reduced capital and operational costs due to the operation at high cell densities (HCD) and improved productivities. HCDs are achieved by cell retention and continuous medium exchange, which is often based on the cell-specific perfusion rate (CSPR). To obtain a cost-productive process the perfusion rate must be determined for each process individually. However, determining optimal operating conditions remain labor-intensive and time-consuming experiments, as investigations are performed in lab-scale perfusion bioreactors. Small-scale models such as microwell plates (MWPs) provide an option for screening multiple perfusion rates in parallel in a semi-perfusion mimic. This study investigated two perfusion rate strategies applied to the MWP platform operated in semi-perfusion. The CSPR-based perfusion rate strategy aimed to maintain multiple CSPR values throughout the cultivation and was compared to a cultivation with a perfusion rate of 1 RV d
    Language English
    Publishing date 2024-03-03
    Publishing country United States
    Document type Journal Article
    ZDB-ID 280318-5
    ISSN 1097-0290 ; 0006-3592
    ISSN (online) 1097-0290
    ISSN 0006-3592
    DOI 10.1002/bit.28685
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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  4. Article ; Online: Process intensification strategies toward cell culture-based high-yield production of a fusogenic oncolytic virus.

    Göbel, Sven / Jaén, Karim E / Dorn, Marie / Neumeyer, Victoria / Jordan, Ingo / Sandig, Volker / Reichl, Udo / Altomonte, Jennifer / Genzel, Yvonne

    Biotechnology and bioengineering

    2023  Volume 120, Issue 9, Page(s) 2639–2657

    Abstract: We present a proof-of-concept study for production of a recombinant vesicular stomatitis virus (rVSV)-based fusogenic oncolytic virus (OV), rVSV-Newcastle disease virus (NDV), at high cell densities (HCD). Based on comprehensive experiments in 1 L ... ...

    Abstract We present a proof-of-concept study for production of a recombinant vesicular stomatitis virus (rVSV)-based fusogenic oncolytic virus (OV), rVSV-Newcastle disease virus (NDV), at high cell densities (HCD). Based on comprehensive experiments in 1 L stirred tank reactors (STRs) in batch mode, first optimization studies at HCD were carried out in semi-perfusion in small-scale cultivations using shake flasks. Further, a perfusion process was established using an acoustic settler for cell retention. Growth, production yields, and process-related impurities were evaluated for three candidate cell lines (AGE1.CR, BHK-21, HEK293SF)infected at densities ranging from 15 to 30 × 10
    MeSH term(s) Animals ; Oncolytic Viruses/genetics ; Cell Culture Techniques ; Bioreactors ; Cell Line ; Vesiculovirus/genetics ; Virus Cultivation
    Language English
    Publishing date 2023-02-21
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 280318-5
    ISSN 1097-0290 ; 0006-3592
    ISSN (online) 1097-0290
    ISSN 0006-3592
    DOI 10.1002/bit.28353
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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