Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations
Monoclonal antibodies (mAb) and other biological drugs are affected by enzymatic polysorbate (PS) degradation that reduces product stability and jeopardizes the supply of innovative medicines. PS represents a critical surfactant stabilizing the active pharmaceutical ingredients, which are produced b...
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| Language: | English |
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Taylor & Francis Group
2024-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2024.2375798 |
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| author | Melanie Maier Linus Weiß Nikolas Zeh Valerie Schmieder-Todtenhaupt Alireza Dehghani Marius Nicolaus Felix Daniel Heinzelmann Benjamin Lindner Moritz Schmidt Joey Studts Patrick Schulz Bernd Reisinger Kerstin Otte Matthias Franzreb Daniel Lakatos Simon Fischer |
| author_facet | Melanie Maier Linus Weiß Nikolas Zeh Valerie Schmieder-Todtenhaupt Alireza Dehghani Marius Nicolaus Felix Daniel Heinzelmann Benjamin Lindner Moritz Schmidt Joey Studts Patrick Schulz Bernd Reisinger Kerstin Otte Matthias Franzreb Daniel Lakatos Simon Fischer |
| author_sort | Melanie Maier |
| collection | DOAJ |
| description | Monoclonal antibodies (mAb) and other biological drugs are affected by enzymatic polysorbate (PS) degradation that reduces product stability and jeopardizes the supply of innovative medicines. PS represents a critical surfactant stabilizing the active pharmaceutical ingredients, which are produced by recombinant Chinese hamster ovary (CHO) cell lines. While the list of potential PS-degrading CHO host cell proteins (HCPs) has grown over the years, tangible data on industrially relevant HCPs are still scarce. By means of a highly sensitive liquid chromatography-tandem mass spectrometry method, we investigated seven different mAb products, resulting in the identification of 12 potentially PS-degrading hydrolases, including the strongly PS-degrading lipoprotein lipase (LPL). Using an LPL knockout CHO host cell line, we were able to stably overexpress and purify the remaining candidate hydrolases through orthogonal affinity chromatography methods, enabling their detailed functional characterization. Applying a PS degradation assay, we found nine mostly secreted, PS-active hydrolases with varying hydrolytic activity. All active hydrolases showed a serine-histidine-aspartate/glutamate catalytical triad. Further, we subjected the active hydrolases to pH-screenings and revealed a diverse range of activity optima, which can facilitate the identification of residual hydrolases during bioprocess development. Ultimately, we compiled our dataset in a risk matrix identifying PAF-AH, LIPA, PPT1, and LPLA2 as highly critical hydrolases based on their cellular expression, detection in purified antibodies, active secretion, and PS degradation activity. With this work, we pave the way toward a comprehensive functional characterization of PS-degrading hydrolases and provide a basis for a future reduction of PS degradation in biopharmaceutical drug products. |
| format | Article |
| id | doaj-art-25ff403b4f7d467a92c72862afa44c17 |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-25ff403b4f7d467a92c72862afa44c172025-01-31T04:19:38ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2375798Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulationsMelanie Maier0Linus Weiß1Nikolas Zeh2Valerie Schmieder-Todtenhaupt3Alireza Dehghani4Marius Nicolaus Felix5Daniel Heinzelmann6Benjamin Lindner7Moritz Schmidt8Joey Studts9Patrick Schulz10Bernd Reisinger11Kerstin Otte12Matthias Franzreb13Daniel Lakatos14Simon Fischer15Bioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyInstitute for Applied Biotechnology, University of Applied Sciences Biberach, Biberach an der Riss, GermanyInstitute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyBioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, GermanyMonoclonal antibodies (mAb) and other biological drugs are affected by enzymatic polysorbate (PS) degradation that reduces product stability and jeopardizes the supply of innovative medicines. PS represents a critical surfactant stabilizing the active pharmaceutical ingredients, which are produced by recombinant Chinese hamster ovary (CHO) cell lines. While the list of potential PS-degrading CHO host cell proteins (HCPs) has grown over the years, tangible data on industrially relevant HCPs are still scarce. By means of a highly sensitive liquid chromatography-tandem mass spectrometry method, we investigated seven different mAb products, resulting in the identification of 12 potentially PS-degrading hydrolases, including the strongly PS-degrading lipoprotein lipase (LPL). Using an LPL knockout CHO host cell line, we were able to stably overexpress and purify the remaining candidate hydrolases through orthogonal affinity chromatography methods, enabling their detailed functional characterization. Applying a PS degradation assay, we found nine mostly secreted, PS-active hydrolases with varying hydrolytic activity. All active hydrolases showed a serine-histidine-aspartate/glutamate catalytical triad. Further, we subjected the active hydrolases to pH-screenings and revealed a diverse range of activity optima, which can facilitate the identification of residual hydrolases during bioprocess development. Ultimately, we compiled our dataset in a risk matrix identifying PAF-AH, LIPA, PPT1, and LPLA2 as highly critical hydrolases based on their cellular expression, detection in purified antibodies, active secretion, and PS degradation activity. With this work, we pave the way toward a comprehensive functional characterization of PS-degrading hydrolases and provide a basis for a future reduction of PS degradation in biopharmaceutical drug products.https://www.tandfonline.com/doi/10.1080/19420862.2024.2375798Polysorbatehydrolasehydrolysisdegradationhost cell proteinlipase |
| spellingShingle | Melanie Maier Linus Weiß Nikolas Zeh Valerie Schmieder-Todtenhaupt Alireza Dehghani Marius Nicolaus Felix Daniel Heinzelmann Benjamin Lindner Moritz Schmidt Joey Studts Patrick Schulz Bernd Reisinger Kerstin Otte Matthias Franzreb Daniel Lakatos Simon Fischer Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations mAbs Polysorbate hydrolase hydrolysis degradation host cell protein lipase |
| title | Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations |
| title_full | Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations |
| title_fullStr | Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations |
| title_full_unstemmed | Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations |
| title_short | Illuminating a biologics development challenge: systematic characterization of CHO cell-derived hydrolases identified in monoclonal antibody formulations |
| title_sort | illuminating a biologics development challenge systematic characterization of cho cell derived hydrolases identified in monoclonal antibody formulations |
| topic | Polysorbate hydrolase hydrolysis degradation host cell protein lipase |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2375798 |
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