Custom made inclusion bodies: impact of classical process parameters and physiological parameters on inclusion body quality attributes

Microbial Cell Factories
Christoph SloukaChristoph Herwig

Abstract

The bacterium E. coli is a major host for recombinant protein production of non-glycosylated products. Depending on the expression strategy, the recombinant protein can be located intracellularly. In many cases the formation of inclusion bodies (IBs), protein aggregates inside of the cytoplasm of the cell, is favored in order to achieve high productivities and to cope with toxic products. However, subsequent downstream processing, including homogenization of the cells, centrifugation or solubilization of the IBs, is prone to variable process performance or can be characterized by low extraction yields as published elsewhere. It is hypothesized that variations in IB quality attributes (QA) are responsible for those effects and that such attributes can be controlled by upstream process conditions. This contribution is aimed at analyzing how standard process parameters, such as pH and temperature (T) as well as different controlled levels of physiological parameters, such as specific substrate uptake rates, can vary IB quality attributes. Classical process parameters like pH and T influence the expression of analyzed IB. The effect on the three QAs titer, size and purity could be successfully revealed. The developed data driven mo...Continue Reading

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Citations

Jul 4, 2019·Bioprocess and Biosystems Engineering·Alexander PekarskyOliver Spadiut
Jul 3, 2020·International Journal of Molecular Sciences·Diana HumerOliver Spadiut
Feb 12, 2020·Advanced Science·Julieta M SánchezAntonio Villaverde
Nov 19, 2019·Frontiers in Bioengineering and Biotechnology·Julian KoppChristoph Slouka
Jul 29, 2020·Frontiers in Bioengineering and Biotechnology·Garima ThakurAnurag S Rathore
Dec 21, 2018·Applied Microbiology and Biotechnology·Christoph SloukaChristoph Herwig
Dec 12, 2019·Frontiers in Bioengineering and Biotechnology·Julian KoppChristoph Herwig
Jul 13, 2020·Microbial Cell Factories·Elena KrachmarovaGenoveva Nacheva
Nov 27, 2020·Frontiers in Bioengineering and Biotechnology·Julian KoppDavid J Wurm
Jan 20, 2020·Trends in Biotechnology·Priyank SinghviAmulya K Panda
Jun 3, 2021·Scientific Reports·Stefan KittlerJulian Kopp
Jan 1, 2020·Journal of Biotechnology·Britta EggenreichOliver Spadiut

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Methods Mentioned

BETA
glycosylation
FCS
AFM
transmission electron microscopy

Software Mentioned

Fiji
OriginPro
DeNovo
SDS
control
DasGip
FCS Express
CyView
DASware
ImageJ

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