Abstract
To explore the feasability of larch (Larix decidua Mill.) embryogenic cell culture as alternative plant expression system, protein stability in fresh and conditioned medium was characterized in this study and compared to tobacco BY2 suspension culture. Fresh and conditioned media were spiked with 1 μg human IgG and IgG content was determined by ELISA after 24 h incubation. In fresh media, IgG recovery rate decreased to 12–23%. Adsorption on vessel walls probably is the best explanation for this IgG loss and EDTA in the medium strongly influenced wall adsorption. A high IgG recovery rate occurred in all conditioned cell culture media (7 or 14 days after inoculation). Changes in the low molecular weight-constitution of conditioned medium, rather than co-secreted polymers, are responsible for IgG stability in the cell suspension cultures.
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Acknowledgements
The authors wish to thank S.Schillberg, Fraunhofer Institute of Molecular Biology and Applied Ecology, Aachen, for provision of BY2 cell cultures and K.-P. Linscheid for helpful discussions. Finally, we thank M. Becker and H. Peisker for excellent technical assistance.
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Calinski, A., Classen, B., Zoglauer, K. et al. IgG stability in fresh and conditioned medium of tobacco (Nicotiana tabacum) and larch (Larix decidua) embryogenic suspension cultures. Biotechnol Lett 31, 771–778 (2009). https://doi.org/10.1007/s10529-009-9920-9
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DOI: https://doi.org/10.1007/s10529-009-9920-9