Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid
Research background. Protein A affinity chromatography is a well-established method currently used in the pharmaceutical industry. However, the high costs usually associated with chromatographic separation of protein A and the difficulties in continuous operation make the investigation of alternativ...
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| Format: | Article |
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University of Zagreb Faculty of Food Technology and Biotechnology
2023-01-01
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| Series: | Food Technology and Biotechnology |
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| Online Access: | https://hrcak.srce.hr/file/447116 |
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| author | Dorottya Vaskó Júlia Domján Bence Szűcs László Bakk Péter Hajdinák György Marosi Zsombor K. Nagy Edit Hirsch Csaba Fehér |
| author_facet | Dorottya Vaskó Júlia Domján Bence Szűcs László Bakk Péter Hajdinák György Marosi Zsombor K. Nagy Edit Hirsch Csaba Fehér |
| author_sort | Dorottya Vaskó |
| collection | DOAJ |
| description | Research background. Protein A affinity chromatography is a well-established method currently used in the pharmaceutical industry. However, the high costs usually associated with chromatographic separation of protein A and the difficulties in continuous operation make the investigation of alternative purification methods very important.
Experimental approach. In this study, extraction/back-extraction and precipitation/dissolution methods were developed and optimised. They were compared with protein A and cation exchange chromatography separations in terms of yield of monoclonal antibody (mAb) and amount of residual impurities, such as DNA and host cell proteins, and amount of mAb aggregates. For a comprehensive comparison of the different methods, experiments were carried out with the same cell-free fermentation broth containing adalimumab.
Results and conclusions. Protein A and cation exchange chromatographic separations resulted in high yield and purity of adalimumab. The precipitation-based process resulted in high yield but with lower purity. The extraction-based purification resulted in low yield and purity. Thus, the precipitation-based method proved to be more promising than the extraction-based method for direct purification of adalimumab from harvested cell culture fluid.
Novelty and scientific contribution. Although alternative purification methods may offer the advantages of simplicity and low-cost operation, further significant improvements are required to compete with the performance of chromatographic separations of adalimumab from true fermentation broth. |
| first_indexed | 2024-04-24T09:05:29Z |
| format | Article |
| id | doaj.art-c6272988b86f4b37873d6086ab3a3fa4 |
| institution | Directory Open Access Journal |
| issn | 1330-9862 1334-2606 |
| language | English |
| last_indexed | 2024-04-24T09:05:29Z |
| publishDate | 2023-01-01 |
| publisher | University of Zagreb Faculty of Food Technology and Biotechnology |
| record_format | Article |
| series | Food Technology and Biotechnology |
| spelling | doaj.art-c6272988b86f4b37873d6086ab3a3fa42024-04-15T19:02:16ZengUniversity of Zagreb Faculty of Food Technology and BiotechnologyFood Technology and Biotechnology1330-98621334-26062023-01-0161333934910.17113/ftb.61.03.23.8094Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture FluidDorottya Vaskó0Júlia Domján1Bence Szűcs2László Bakk3Péter Hajdinák4György Marosi5Zsombor K. Nagy6Edit Hirsch7Csaba Fehér8Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryDepartment of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, 1111 Budapest, HungaryResearch background. Protein A affinity chromatography is a well-established method currently used in the pharmaceutical industry. However, the high costs usually associated with chromatographic separation of protein A and the difficulties in continuous operation make the investigation of alternative purification methods very important. Experimental approach. In this study, extraction/back-extraction and precipitation/dissolution methods were developed and optimised. They were compared with protein A and cation exchange chromatography separations in terms of yield of monoclonal antibody (mAb) and amount of residual impurities, such as DNA and host cell proteins, and amount of mAb aggregates. For a comprehensive comparison of the different methods, experiments were carried out with the same cell-free fermentation broth containing adalimumab. Results and conclusions. Protein A and cation exchange chromatographic separations resulted in high yield and purity of adalimumab. The precipitation-based process resulted in high yield but with lower purity. The extraction-based purification resulted in low yield and purity. Thus, the precipitation-based method proved to be more promising than the extraction-based method for direct purification of adalimumab from harvested cell culture fluid. Novelty and scientific contribution. Although alternative purification methods may offer the advantages of simplicity and low-cost operation, further significant improvements are required to compete with the performance of chromatographic separations of adalimumab from true fermentation broth.https://hrcak.srce.hr/file/447116monoclonal antibodypurification technologyprotein A chromatographyprecipitationextraction |
| spellingShingle | Dorottya Vaskó Júlia Domján Bence Szűcs László Bakk Péter Hajdinák György Marosi Zsombor K. Nagy Edit Hirsch Csaba Fehér Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid Food Technology and Biotechnology monoclonal antibody purification technology protein A chromatography precipitation extraction |
| title | Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid |
| title_full | Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid |
| title_fullStr | Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid |
| title_full_unstemmed | Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid |
| title_short | Development and Comparison of Alternative Methods for the Purification of Adalimumab Directly from Harvested Cell Culture Fluid |
| title_sort | development and comparison of alternative methods for the purification of adalimumab directly from harvested cell culture fluid |
| topic | monoclonal antibody purification technology protein A chromatography precipitation extraction |
| url | https://hrcak.srce.hr/file/447116 |
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