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

Title data

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: https://doi.org/10.1002/elsc.201700005

Abstract in another language

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.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: Cell-free expression; CHO cell lysate; Glycosylation; hBMP2; Mammalian cell culture
Institutions of the University: Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Faculties
Result of work at the UBT: Yes
DDC Subjects: 600 Technology, medicine, applied sciences
600 Technology, medicine, applied sciences > 610 Medicine and health
600 Technology, medicine, applied sciences > 620 Engineering
Date Deposited: 20 Jul 2017 11:11
Last Modified: 20 Jul 2017 11:11
URI: https://eref.uni-bayreuth.de/id/eprint/38736