Two-hundred-liter scale fermentation, purification of recombinant human fibroblast growth factor-21, and its anti-diabetic effects on ob/ob mice
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Fibroblast growth factor-21 (FGF-21) is a potential cytokine for type II diabetes mellitus. This study aimed to optimize recombinant human FGF-21 (rhFGF-21) production in Escherichia coli BL21 (DE3) employing high cell density fermentation at a 200-L scale and pilot-scale purification. FGF-21 was eventually expressed in E. coli BL21 (DE3) using human FGF-21 synthetic DNA sequence via the introduction of vector pET-3c; the product is used as seed strain during the fermentation of rhFGF-21. Fermentation of rhFGF-21 was performed in a 30-L and 200-L fermenters. rhFGF-21 was primarily expressed in the form of inclusion bodies after IPTG induction. At the 200-L scale, the bacterial production and expression levels of rhFGF-21 were 38.8 ± 0.6 g/L and 30.9 ± 0.7%, respectively. Additionally, the high purification (98%) of rhFGF-21 was tested with HPLC analysis and reducing & non-reducing SDS-PAGE analysis. The final yield of purified rhFGF-21 was 71.1 ± 13.9 mg/L. The activity of rhFGF-21 stock solution reached at 68.67 ± 8.74 IU/mg. Blood glucose controlling and insulin sensitization were improved with treatment of rhFGF-21 in type II diabetic ob/ob mice. Our results showed that the relatively stable and time-saving pilot-scale production process was successfully established, providing an efficient and cost-effective strategy for large-scale and industrial production of rhFGF-21.
KeywordsFGF-21 Expression Purification Glucose-lowering
This research was funded by a grant from the ministry of science and technology of China (No. 2011ZX09102-004-03), the National Natural Science Foundation of China (No. 81601695), the Natural Science Foundation of Zhejiang Province (No. LY17H150002), and the Wenzhou Science and Technology Agency (No. ZS2017016).
Compliance with ethical standards
All animal experiments were handled in accordance with IACUC guidelines of Wenzhou Medical University (Zhejiang, China), complying with NIH guidelines for the care and use of laboratory animals. This article does not contain any studies with human participants. All authors confirm that ethical principles have been followed in the experiments.
Conflicts of interest
The authors declare that they have no conflict of interest.
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