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Published on: March 29, 2020
PEGylation-aided refolding of globular adiponectin
Mingming Gao1, Yue Tong, Xiangdong Gao
1State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, Tongjiaxiang 24, Nanjing 210009, China.
Globular adiponectin (GAD) refolding for diabetes drug development was improved using polyethylene glycol (PEG) conjugation. This novel PEGylation-aided method efficiently refolded GAD, restoring its glucose-lowering activity in mice.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Development
Background:
- Globular adiponectin (GAD), the active domain of adiponectin, shows potential for anti-diabetic drug development.
- Recombinant GAD production in Escherichia coli often results in inclusion bodies requiring refolding for functionality.
Purpose of the Study:
- To develop an efficient method for refolding GAD using polyethylene glycol (PEG) conjugation.
- To assess the biological activity of refolded GAD.
Main Methods:
- Synthesized a DNA sequence for GAD and constructed an expression plasmid in E. coli.
- Collected and dissolved inclusion bodies in guanidine hydrochloride for PEG conjugation.
- Refolded PEG-conjugated GAD via dialysis and purified using chromatography.
Main Results:
- Developed a novel PEGylation-aided refolding technique for GAD.
- The refolded PEG-conjugated GAD demonstrated significant glucose-lowering activity in mice.
- Confirmed successful refolding and restoration of GAD's biological function.
Conclusions:
- PEGylation-aided refolding is an efficient method for producing functional GAD.
- This technique offers a promising approach for recombinant protein refolding and drug development.
- The method's suitability can be explored for other proteins requiring refolding.
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