de Vries, I.; Schreiber, S.; Boßmann, D.; Hellmann, Z.; Kopatz, J. et al.: Single-use membrane adsorbers for endotoxin removal and purification of endogenous polysialic acid from Escherichia coli K1. In: Biotechnology Reports 17 (2018), S. 110-116. DOI:
https://doi.org/10.1016/j.btre.2018.02.001
Abstract: |
Polysialic acid (polySia) is a promising molecule for various medical applications (e.g., treatment of inflammatory neurodegenerative diseases). In this study a complete production process for human-identical α-(2,8)-linked polySia was developed using a disposable bioreactor for cultivation of Escherichia coli K1 and single-use membrane adsorbers for downstream processing (DSP). The cultivation process was optimized to minimize complex media components and a maturation process after cultivation was established. The maturation led to further product release from the cell surface into the supernatant. Afterwards DSP was established using sodium hydroxide treatment combined with anion exchange membrane adsorbers for endotoxin and DNA depletion. After downstream processing the final product had neither detectable protein nor DNA contamination. Endotoxin content was below 3 EU mg−1. Investigation of the maximal chain length showed no effect of the harsh sodium hydroxide treatment during DSP on the stability of the polySia. Maximal chain length was ∼98 degree of polymerization. © 2018 The Authors
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License of this version: |
CC BY-NC-ND 4.0 Unported - https://creativecommons.org/licenses/by-nc-nd/4.0/
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Publication type: |
Article |
Publishing status: |
publishedVersion |
Publication date: |
2018 |
Keywords english: |
Endotoxin, Escherichia coli K1, Membrane adsorber, Polysialic acid, Single-use, endotoxin, glucose, polysialic acid, sodium hydroxide, anion exchange, Article, bacterial strain, cell disruption, cell maturation, coculture, controlled study, degenerative disease, DNA contamination, DNA determination, downstream processing, dry weight, Escherichia coli, fast protein liquid chromatography, fermentation, flow rate, freeze drying, gel electrophoresis, glucose intake, high performance liquid chromatography, hydrolysis, inflammation, inoculation, membrane, mitochondrial DNA depletion, molecular weight, nonhuman, optical density, pH, polymerase chain reaction, polymerization, priority journal, temperature, ultrafiltration
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DDC: |
660 | Technische Chemie
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