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

 
: Jérôme, V.; Thoring, L.; Salzig, D.; Kubick, S.; Freitag, R.

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Engineering in life sciences 17 (2017), No.10, pp.1097-1107
ISSN: 1618-2863
ISSN: 1618-0240
English
Journal Article, Electronic Publication
Fraunhofer IZI ()

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. Up to 280 ng/mL 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, reached within three 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.

: http://publica.fraunhofer.de/documents/N-464653.html