期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 110, 期 4, 页码 1164-1173出版社
WILEY-BLACKWELL
DOI: 10.1002/bit.24776
关键词
signal peptide; recombinant antibody; high productive; cell-specific productivity; Chinese hamster ovary (CHO); fed batch
资金
- JB Laboratory (MHH)
- Excellence Initiative REBIRTH (Regenerative Biology to Reconstructive Therapy)
- SFB 738 (Optimierung konventioneller and innovativer Transplantate)
- ReGene (Regenerative Medicine and Biology)
- Bundesministerium fur Bildung und Forschung
Recombinant biotherapeutic proteins such as monoclonal antibodies are mostly produced in Chinese hamster ovary (CHO) cells and pharmaceutical companies are interested in an appropriate platform technology for the development of large-scale production processes. A major aim of our study was therefore to improve the secretion efficiency of a recombinant biotherapeutic antibody by optimizing signal peptides. Reporter molecules such as gaussia and vargula luciferase or secreted alkaline phosphatase are frequently used to this end. In striking contrast, we used a biotherapeutic antibody that was fused to 16 different signal peptides during our study. In this way, the secretion efficiency of the recombinant antibody has been analyzed by transient expression experiments in CHO cell lines. Compared to the control signal peptide, it was not possible to achieve higher efficiencies with signal peptides derived from a variety of species or even natural immunoglobulin G signal peptides. The best results were obtained with natural signal peptides derived from human albumin and human azurocidin. These results were confirmed by fed-batch experiments with stably transfected cell pools, in which cell-specific productivities up to 90pgcell1day1 and product concentrations up to 4gL1 could be determined using the albumin signal peptide. Finally, the applicability of the identified signal peptides for both different antibodies and non-antibody products was demonstrated by transient expression experiments. In conclusion, it was found that signal peptides derived from human albumin and human azurocidin are most appropriate to generate cell lines with clearly improved production rates suitable for commercial purposes in a product-independent manner. Biotechnol. Bioeng. 2013; 110: 11641173. (c) 2012 Wiley Periodicals, Inc.
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