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 -