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

作者：Jerome Valerie; Thoring Lena; Salzig Denise; Kubick Stefan; Freitag Ruth^*

来源：Engineering in Life Sciences, 2017, 17(10): 1097-1107.

DOI：10.1002/elsc.201700005

• Cell-free expression
• CHO cell lysate
• Glycosylation
• hBMP2
• Mammalian cell culture

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 153pg/mL. Up to 280ng/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 40g/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.

• 出版日期2017-10