A high density CHO-S transient transfection system: Comparison of ExpiCHO and Expi293
| Autor: | Fang Sun, Susan Barkowski-Clark, Rekha Patel, Tadas Panavas, Nina K. Jain, Richard Altman, Voegtli Walter C, Alinafe Mpanda, Jian Liu, Alyssa Neill, Jonathan Zmuda, Krista Johnson, Henry Chiou, Adriana Kita, Robert Ballinger, Ryan Schulz, Brian P. Huta, Chao Yan Liu |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Glycosylation Gene Expression CHO Cells Transfection 01 natural sciences 03 medical and health sciences chemistry.chemical_compound Cricetulus 010608 biotechnology Cricetinae Protein biosynthesis Animals Humans biology Chinese hamster ovary cell HEK 293 cells Embryonic stem cell Recombinant Proteins Titer 030104 developmental biology HEK293 Cells chemistry Biochemistry Cell culture biology.protein Antibody Protein Processing Post-Translational Biotechnology |
| Zdroj: | Protein expression and purification. 134 |
| ISSN: | 1096-0279 |
| Popis: | Chinese Hamster Ovary (CHO) cells are the principal mammalian host used for stable cell line generation and biotherapeutic protein production. Until recently, production of milligrams to grams of protein in CHO transient systems was challenging. As such, Human Embryonic Kidney (HEK293) cells are the most common mammalian cell type used for transient transfection. The post-translational modifications (PTMs) of a protein are dictated in part by the cell line used for expression, and changes in PTMs have been shown to affect both the activity and biophysical properties of proteins. Therefore, it is potentially advantageous to keep the host cell type consistent throughout drug discovery and development. To this end, we compared the ExpiCHO system, a high density CHO-S transient transfection system, to the Expi293 and FreeStyle MAX CHO transient systems. Fourteen proteins were expressed in both the Expi293 and ExpiCHO systems. For a majority of proteins tested, the protein titers observed with the ExpiCHO system were higher than those seen with both the FreeStyle MAX CHO and Expi293 systems. Antibodies expressed using the ExpiCHO system had glycosylation patterns more similar to antibodies produced in stable CHO cell lines than Expi293-derived antibodies. However, culture duration and temperature were found to affect protein titer, monodispersity, enzyme activity, and PTMs and should be carefully selected when using the ExpiCHO system. The ExpiCHO transient transfection systems allows for facile production of milligrams to grams of protein in CHO cells and de-risks the transition from transient to stable material during drug development. |
| Databáze: | OpenAIRE |
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