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| Expression of Soluble Anti-TNF-α Fab in E.coli: Optimization for Technological Process |
| ZHANG Yu-meng1,2, TONG Mei2, LU Xiao-dong2, MI Yue1, MO Ting1,2, LIU Jin-yi2, YAO Wen-bing1 |
1 Institute of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China;
2 Beijing Tri-Prime Gene Pharmaceutical Co. Ltd., Beijing 102600, China |
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Abstract Subject:The anti-TNF-α Fab recombinant plasmid to achieve an efficient soluble expression of anti-TNF-α Fab in periplasmic space of E. coli, and to improve the yield of the target protein by optimizing technological process were constructed. Method:After fermentation, extraction method was optimized for a higher Fab yield by DOE to extract the target protein of periplasm. Ion exchange chromatography (IEX) was applied to clear the extraction, and purification was achieved by hydrophobic interaction chromatography (HIC) and Protein L affinity chromatography. Results:An optimal extraction condition by DOE and the yield was up to 23.5mg/L, the overall recovery rate was above 24% and the purity was above 90%. Ion exchange chromatography provided a good protection for affinity column. The result of affinity assay indicated that the anti-TNF-α Fab had excellent bioactivities. Conclusion:A foundation for further amplification of the production by optimizing extraction and purification process have been made.
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Received: 09 March 2016
Published: 25 September 2016
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