Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells
(1) Recombinant protein production in mammalian cells is either based on transient transfection processes, often inefficient and underlying high batch-to-batch variability, or on laborious generation of stable cell lines. Alternatively, BacMam, a transduction process using the baculovirus, can be em...
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| Format: | Article |
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MDPI AG
2024-03-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/16/3/426 |
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| author | Maria Toth Manuel Reithofer Gregory Dutra Patricia Pereira Aguilar Astrid Dürauer Reingard Grabherr |
| author_facet | Maria Toth Manuel Reithofer Gregory Dutra Patricia Pereira Aguilar Astrid Dürauer Reingard Grabherr |
| author_sort | Maria Toth |
| collection | DOAJ |
| description | (1) Recombinant protein production in mammalian cells is either based on transient transfection processes, often inefficient and underlying high batch-to-batch variability, or on laborious generation of stable cell lines. Alternatively, BacMam, a transduction process using the baculovirus, can be employed. (2) Six transfecting agents were compared to baculovirus transduction in terms of transient and stable protein expression characteristics of the model protein ACE2-eGFP using HEK293-6E, CHO-K1, and Vero cell lines. Furthermore, process optimization such as expression enhancement using sodium butyrate and TSA or baculovirus purification was assessed. (3) Baculovirus transduction efficiency was superior to all transfection agents for all cell lines. Transduced protein expression was moderate, but an 18-fold expression increase was achieved using the enhancer sodium butyrate. Ultracentrifugation of baculovirus from a 3.5 L bioreactor significantly improved the transduction efficiency and protein expression. Stable cell lines were obtained with each baculovirus transduction, yet stable cell line generation after transfection was highly unreliable. (4) This study demonstrated the superiority of the BacMam platform to standard transfections. The baculovirus efficiently transduced an array of cell lines both transiently and stably and achieved the highest efficiency for all tested cell lines. The feasibility of the scale-up of baculovirus production was demonstrated and the possibility of baculovirus purification was successfully explored. |
| first_indexed | 2024-04-24T17:46:15Z |
| format | Article |
| id | doaj.art-824026465af14447a8c9b3ea9ac786a0 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-04-24T17:46:15Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-824026465af14447a8c9b3ea9ac786a02024-03-27T14:07:50ZengMDPI AGViruses1999-49152024-03-0116342610.3390/v16030426Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian CellsMaria Toth0Manuel Reithofer1Gregory Dutra2Patricia Pereira Aguilar3Astrid Dürauer4Reingard Grabherr5Department of Biotechnology, University of Natural Resources and Life Sciences Vienna, 1190 Vienna, AT, AustriaDepartment of Biotechnology, University of Natural Resources and Life Sciences Vienna, 1190 Vienna, AT, AustriaAustrian Centre of Industrial Biotechnology (ACIB), 1190 Vienna, AT, AustriaAustrian Centre of Industrial Biotechnology (ACIB), 1190 Vienna, AT, AustriaDepartment of Biotechnology, University of Natural Resources and Life Sciences Vienna, 1190 Vienna, AT, AustriaDepartment of Biotechnology, University of Natural Resources and Life Sciences Vienna, 1190 Vienna, AT, Austria(1) Recombinant protein production in mammalian cells is either based on transient transfection processes, often inefficient and underlying high batch-to-batch variability, or on laborious generation of stable cell lines. Alternatively, BacMam, a transduction process using the baculovirus, can be employed. (2) Six transfecting agents were compared to baculovirus transduction in terms of transient and stable protein expression characteristics of the model protein ACE2-eGFP using HEK293-6E, CHO-K1, and Vero cell lines. Furthermore, process optimization such as expression enhancement using sodium butyrate and TSA or baculovirus purification was assessed. (3) Baculovirus transduction efficiency was superior to all transfection agents for all cell lines. Transduced protein expression was moderate, but an 18-fold expression increase was achieved using the enhancer sodium butyrate. Ultracentrifugation of baculovirus from a 3.5 L bioreactor significantly improved the transduction efficiency and protein expression. Stable cell lines were obtained with each baculovirus transduction, yet stable cell line generation after transfection was highly unreliable. (4) This study demonstrated the superiority of the BacMam platform to standard transfections. The baculovirus efficiently transduced an array of cell lines both transiently and stably and achieved the highest efficiency for all tested cell lines. The feasibility of the scale-up of baculovirus production was demonstrated and the possibility of baculovirus purification was successfully explored.https://www.mdpi.com/1999-4915/16/3/426BacMambaculovirustransfectiontransductionprotein productionexpression enhancement |
| spellingShingle | Maria Toth Manuel Reithofer Gregory Dutra Patricia Pereira Aguilar Astrid Dürauer Reingard Grabherr Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells Viruses BacMam baculovirus transfection transduction protein production expression enhancement |
| title | Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells |
| title_full | Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells |
| title_fullStr | Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells |
| title_full_unstemmed | Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells |
| title_short | Comprehensive Comparison of Baculoviral and Plasmid Gene Delivery in Mammalian Cells |
| title_sort | comprehensive comparison of baculoviral and plasmid gene delivery in mammalian cells |
| topic | BacMam baculovirus transfection transduction protein production expression enhancement |
| url | https://www.mdpi.com/1999-4915/16/3/426 |
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