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Comparison of cell-based vs. cell-free mammalian systems for the production of a recombinant human bone morphogenic growth factor

Titelangaben

Jérôme, Valérie ; Thoring, Lena ; Salzig, Denise ; Kubick, Stefan ; Freitag, Ruth:
Comparison of cell-based vs. cell-free mammalian systems for the production of a recombinant human bone morphogenic growth factor.
In: Engineering in Life Sciences. (2017) .
ISSN 1618-2863
DOI: 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: 20 Jul 2017 11:11
URI: https://eref.uni-bayreuth.de/id/eprint/38736