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Jérôme, Valérie ; Thoring, Lena ; Salzig, Denise ; Kubick, Stefan ; Freitag, Ruth:
Comparison of cell-based versus cell-free mammalian systems for the production of a recombinant human bone morphogenic growth factor.
In: Engineering in Life Sciences.
Bd. 17
(2017)
Heft 10
.
- S. 1097-1107.
ISSN 1618-2863
DOI: https://doi.org/10.1002/elsc.201700005
Abstract
The human bone morphogenetic protein-2 (hBMP2) is a glycoprotein, which induces de novo bone formation. Here, recombinant production in stably transfected Chinese Hamster Ovary (CHO) cells is compared to transient expression in Human Embryo Kidney (HEK) cells and cell-free synthesis in CHO cell lysates containing microsomal structures as sites of post-translational processing. In case of the stably transfected cells, growth rates and viabilities were similar to those of the parent cells, while entry into the death phase of the culture was delayed. The maximum achievable rhBMP2 concentration in these cultures was 153 pg mL−1. Up to 280 ng mL−1 could be produced in the transient expression system. In both cases the rhBMP-2 was found to interact with the producer cells, which presumably contributed to the low yields. In the cell-free system, hBMP2 yields could be increased to almost 40 μg mL−1, reached within 3 hours. The cell-free system thus approached productivities for the active (renatured) protein previously only recorded for bacterial hosts, while assuring comprehensive post-translational processing.
Weitere Angaben
| Publikationsform: | Artikel in einer Zeitschrift |
|---|---|
| Begutachteter Beitrag: | Ja |
| Keywords: | Cell-free expression; CHO cell lysate; Glycosylation; hBMP2; Mammalian cell culture |
| Institutionen der Universität: | Fakultäten > Fakultät für Ingenieurwissenschaften Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik Fakultäten |
| Titel an der UBT entstanden: | Ja |
| Themengebiete aus DDC: | 600 Technik, Medizin, angewandte Wissenschaften 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit 600 Technik, Medizin, angewandte Wissenschaften > 620 Ingenieurwissenschaften |
| Eingestellt am: | 20 Jul 2017 11:11 |
| Letzte Änderung: | 09 Nov 2022 08:06 |
| URI: | https://eref.uni-bayreuth.de/id/eprint/38736 |