Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture
Abstract Continuous precipitation coupled with continuous tangential flow filtration is a cost‐effective alternative for the capture of recombinant antibodies from crude cell culture supernatant. The removal of surge tanks between unit operations, by the adoption of tubular reactors, maintains a con...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley-VCH
2023-10-01
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| Series: | Engineering in Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/elsc.202300219 |
| _version_ | 1797667875784228864 |
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| author | Gabriele Recanati Magdalena Pappenreiter Christoph Gstoettner Patrick Scheidl Elena Domínguez Vega Bernhard Sissolak Alois Jungbauer |
| author_facet | Gabriele Recanati Magdalena Pappenreiter Christoph Gstoettner Patrick Scheidl Elena Domínguez Vega Bernhard Sissolak Alois Jungbauer |
| author_sort | Gabriele Recanati |
| collection | DOAJ |
| description | Abstract Continuous precipitation coupled with continuous tangential flow filtration is a cost‐effective alternative for the capture of recombinant antibodies from crude cell culture supernatant. The removal of surge tanks between unit operations, by the adoption of tubular reactors, maintains a continuous harvest and mass flow of product with the advantage of a narrow residence time distribution (RTD). We developed a continuous process implementing two orthogonal precipitation methods, CaCl2 precipitation for removal of host‐cell DNA and polyethylene glycol (PEG) for capturing the recombinant antibody, with no influence on the glycosylation profile. Our lab‐scale prototype consisting of two tubular reactors and two stages of tangential flow microfiltration was continuously operated for up to 8 days in a truly continuous fashion and without any product flow interruption, both as a stand‐alone capture and as an integrated perfusion‐capture. Furthermore, we explored the use of a negatively charged membrane adsorber for flow‐through anion exchange as first polishing step. We obtained a product recovery of approximately 80% and constant product quality, with more than two logarithmic reduction values (LRVs) for both host‐cell proteins and host‐cell DNA by the combination of the precipitation‐based capture and the first polishing step. |
| first_indexed | 2024-03-11T20:20:54Z |
| format | Article |
| id | doaj.art-eea721d339984f26a23f3ab7209c98dd |
| institution | Directory Open Access Journal |
| issn | 1618-0240 1618-2863 |
| language | English |
| last_indexed | 2024-03-11T20:20:54Z |
| publishDate | 2023-10-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj.art-eea721d339984f26a23f3ab7209c98dd2023-10-03T07:06:04ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632023-10-012310n/an/a10.1002/elsc.202300219Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody captureGabriele Recanati0Magdalena Pappenreiter1Christoph Gstoettner2Patrick Scheidl3Elena Domínguez Vega4Bernhard Sissolak5Alois Jungbauer6Department of Biotechnology University of Natural Resources and Life Sciences Vienna AustriaDepartment of Biotechnology University of Natural Resources and Life Sciences Vienna AustriaCenter for Proteomics and Metabolomics Leiden University Medical Center Leiden The NetherlandsDepartment of Biotechnology University of Natural Resources and Life Sciences Vienna AustriaCenter for Proteomics and Metabolomics Leiden University Medical Center Leiden The NetherlandsCenter for Proteomics and Metabolomics Leiden University Medical Center Leiden The NetherlandsDepartment of Biotechnology University of Natural Resources and Life Sciences Vienna AustriaAbstract Continuous precipitation coupled with continuous tangential flow filtration is a cost‐effective alternative for the capture of recombinant antibodies from crude cell culture supernatant. The removal of surge tanks between unit operations, by the adoption of tubular reactors, maintains a continuous harvest and mass flow of product with the advantage of a narrow residence time distribution (RTD). We developed a continuous process implementing two orthogonal precipitation methods, CaCl2 precipitation for removal of host‐cell DNA and polyethylene glycol (PEG) for capturing the recombinant antibody, with no influence on the glycosylation profile. Our lab‐scale prototype consisting of two tubular reactors and two stages of tangential flow microfiltration was continuously operated for up to 8 days in a truly continuous fashion and without any product flow interruption, both as a stand‐alone capture and as an integrated perfusion‐capture. Furthermore, we explored the use of a negatively charged membrane adsorber for flow‐through anion exchange as first polishing step. We obtained a product recovery of approximately 80% and constant product quality, with more than two logarithmic reduction values (LRVs) for both host‐cell proteins and host‐cell DNA by the combination of the precipitation‐based capture and the first polishing step.https://doi.org/10.1002/elsc.202300219cross flow filtrationIgGintegrated processingprecipitation |
| spellingShingle | Gabriele Recanati Magdalena Pappenreiter Christoph Gstoettner Patrick Scheidl Elena Domínguez Vega Bernhard Sissolak Alois Jungbauer Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture Engineering in Life Sciences cross flow filtration IgG integrated processing precipitation |
| title | Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture |
| title_full | Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture |
| title_fullStr | Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture |
| title_full_unstemmed | Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture |
| title_short | Integration of a perfusion reactor and continuous precipitation in an entirely membrane‐based process for antibody capture |
| title_sort | integration of a perfusion reactor and continuous precipitation in an entirely membrane based process for antibody capture |
| topic | cross flow filtration IgG integrated processing precipitation |
| url | https://doi.org/10.1002/elsc.202300219 |
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