دورية أكاديمية
A high density CHO-S transient transfection system: Comparison of ExpiCHO and Expi293.
| العنوان: | A high density CHO-S transient transfection system: Comparison of ExpiCHO and Expi293. |
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| المؤلفون: | Jain NK; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Barkowski-Clark S; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Altman R; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Johnson K; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Sun F; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Zmuda J; Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States., Liu CY; Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States., Kita A; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Schulz R; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Neill A; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Ballinger R; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Patel R; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Liu J; Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States., Mpanda A; Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States., Huta B; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Chiou H; Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States., Voegtli W; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States., Panavas T; Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States. Electronic address: Tadas.Panavas@alexion.com. |
| المصدر: | Protein expression and purification [Protein Expr Purif] 2017 Jun; Vol. 134, pp. 38-46. Date of Electronic Publication: 2017 Mar 23. |
| نوع المنشور: | Comparative Study; Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 9101496 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0279 (Electronic) Linking ISSN: 10465928 NLM ISO Abbreviation: Protein Expr Purif Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Orlando, FL : Academic Press Original Publication: San Diego : Academic Press, c1990- |
| مواضيع طبية MeSH: | Gene Expression*, Transfection/*methods, Animals ; CHO Cells ; Cricetinae ; Cricetulus ; HEK293 Cells ; Humans ; Protein Processing, Post-Translational ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics |
| مستخلص: | 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. (Copyright © 2017 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: CHO cells; Glycosylation; HEK293 cells; Post-translational modification; Transient transfection |
| المشرفين على المادة: | 0 (Recombinant Proteins) |
| تواريخ الأحداث: | Date Created: 20170327 Date Completed: 20170807 Latest Revision: 20170807 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.pep.2017.03.018 |
| PMID: | 28342833 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1096-0279 |
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| DOI: | 10.1016/j.pep.2017.03.018 |