Details
| Original language | English |
|---|---|
| Pages (from-to) | 110-116 |
| Number of pages | 7 |
| Journal | Biotechnology Reports |
| Volume | 17 |
| Early online date | 8 Feb 2018 |
| Publication status | Published - Mar 2018 |
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.
Keywords
- Endotoxin, Escherichia coli K1, Membrane adsorber, Polysialic acid, Single-use
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
Sustainable Development Goals
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In: Biotechnology Reports, Vol. 17, 03.2018, p. 110-116.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Single-use membrane adsorbers for endotoxin removal and purification of endogenous polysialic acid from Escherichia coli K1
AU - de Vries, Ingo
AU - Schreiber, Sarah
AU - Boßmann, Daniel
AU - Kopatz, Jens
AU - Neumann, Harald
AU - Beutel, Sascha
N1 - Funding information: This study was funded by the German Research Foundation (DFG, grant numbers: DFG-NE507/14-1 and DFG-SCHE279/35-1 ) and the German Federal Ministry of Education and Research (BMBF, grant numbers: BMBF-03VP00271 and BMBF-03VP00273 ). We thank Dr. Rita Gerardy-Schahn (Hannover Medical School) for generously providing the bacterial strain for the polysialic acid production. Furthermore, we acknowledge all associates for helpful discussions and technical support. The publication of this article was funded by the Open Access Fund of the Leibniz Universität Hannover .
PY - 2018/3
Y1 - 2018/3
N2 - 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.
AB - 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.
KW - Endotoxin
KW - Escherichia coli K1
KW - Membrane adsorber
KW - Polysialic acid
KW - Single-use
UR - http://www.scopus.com/inward/record.url?scp=85041928992&partnerID=8YFLogxK
U2 - 10.1016/j.btre.2018.02.001
DO - 10.1016/j.btre.2018.02.001
M3 - Article
VL - 17
SP - 110
EP - 116
JO - Biotechnology Reports
JF - Biotechnology Reports
ER -