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Published on: December 12, 2017
Solubilization and refolding with simultaneous purification of recombinant human stem cell factor
Chaozhan Wang1, Jiahua Liu, Lili Wang
1Institute of Modern Separation Science, Department of Chemistry, Northwest University, 229 Tai Bai North Road, Xi'an 710069, People's Republic of China. czwang@nwu.edu.cn
Applied Biochemistry and Biotechnology
|May 6, 2008
Summary
Recombinant human stem cell factor (rhSCF) solubilization and refolding were optimized using high pH and low urea concentrations. Ion exchange chromatography (IEC) proved most effective for large-scale rhSCF production.
Area of Science:
- Biotechnology
- Protein Chemistry
- Bioprocessing
Background:
- Recombinant human stem cell factor (rhSCF) is crucial for cell growth.
- Inclusion bodies from Escherichia coli expression pose challenges for rhSCF production.
Purpose of the Study:
- To optimize the solubilization and refolding of rhSCF inclusion bodies.
- To evaluate the efficiency of different refolding and purification methods.
Main Methods:
- Investigated the effects of pH and urea concentration on rhSCF inclusion body solubilization.
- Employed ion exchange chromatography (IEC) for refolding and purification.
- Compared IEC refolding with dilution and dialysis refolding methods.
Main Results:
- High pH solutions with low urea concentrations significantly improved rhSCF solubilization.
- IEC achieved high purity (96.3%) and specific activity (7.8 x 10^5 IU/mg) with 43.0% mass recovery.
- IEC refolding was more efficient than dilution or dialysis methods.
Conclusions:
- Optimized conditions (high pH, low urea) facilitate rhSCF solubilization and renaturation.
- IEC is an efficient method for refolding and purifying rhSCF.
- The developed method shows potential for large-scale rhSCF production.
