Jérôme Valérie, Thoring Lena, Salzig Denise, Kubick Stefan, Freitag Ruth
Chair for Process Biotechnology University of Bayreuth Germany.
Department of Cell-free and Cell-based Bioproduction, Fraunhofer Institute for Cell Therapy and Immunology (IZI) Branch Bioanalytics and Bioprocesses Potsdam-Golm (IZI-BB) Germany.
Eng Life Sci. 2017 Aug 7;17(10):1097-1107. doi: 10.1002/elsc.201700005. eCollection 2017 Oct.
The human bone morphogenetic protein-2 (hBMP2) is a glycoprotein, which induces 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.
人骨形态发生蛋白-2（hBMP2）是一种诱导骨形成的糖蛋白。在此，将稳定转染的中国仓鼠卵巢（CHO）细胞中的重组生产与人类胚胎肾（HEK）细胞中的瞬时表达以及含有微粒体结构作为翻译后加工位点的CHO细胞裂解物中的无细胞合成进行比较。对于稳定转染的细胞，其生长速率和活力与亲本细胞相似，而培养进入死亡期则延迟。这些培养物中可达到的重组人骨形态发生蛋白-2（rhBMP2）的最大浓度为153 pg/mL。在瞬时表达系统中可产生高达280 ng/mL的rhBMP2。在这两种情况下，均发现rhBMP-2与生产细胞相互作用，这可能是产量低的原因。在无细胞系统中，hBMP2产量可在三小时内提高到近40 μg/mL。因此，无细胞系统接近了之前仅在细菌宿主中记录的活性（复性）蛋白的生产率，同时确保了全面的翻译后加工。
