High pressure homogenization is a key unit operation in inclusion body processing
Britta Eggenreich1, David Johannes Wurm1, Vignesh Rajamanickam1
1Institute of Chemical, Environmental and Bioscience Engineering, Research Group Integrated Bioprocess Development, Technische Universität Wien, Gumpendorferstrasse 1a, 1060 Vienna, Austria.
Journal of Biotechnology
|June 22, 2021
Summary
Recombinant protein production using inclusion bodies (IBs) in E. coli can be efficient. Optimizing homogenization, washing, and solubilization processes significantly improves protein yield, purity, and biological activity.
Area of Science:
- Biotechnology
- Protein Engineering
- Bioprocessing
Background:
- Recombinant protein production in E. coli frequently results in inclusion bodies (IBs).
- Despite processing challenges, IBs offer advantages like high purity and potential for correct protein folding.
- Evaluating IB processes involves assessing product yield, purity, and biological activity post-refolding.
Purpose of the Study:
- To develop an integrated refolding process for recombinant proteins from E. coli inclusion bodies.
- To analyze the effects and interdependencies of homogenization, IB wash, and IB solubilization unit operations.
Main Methods:
- Investigated the impact of homogenization, IB wash, and IB solubilization on refolding outcomes.
- Analyzed interdependencies between these unit operations to identify synergistic effects.
Main Results:
- Identified significant interactions between homogenization and IB wash, and homogenization and solubilization.
- Homogenization emerged as a critical unit operation, influencing yield (2-fold), purity (1.2-fold), and biological activity (2.5-fold).
Conclusions:
- An integrated approach is crucial for optimizing IB refolding processes.
- Homogenization conditions are key determinants of refolded protein quality and yield.
Keywords:
High pressure homogenizationInclusion body processingInclusion body sizeProduct purityRefolding yield
