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Preparation and PEGylation of exendin-4 peptide secreted from yeast Pichia pastoris
Zhou, J; Cai, ZH; Li, L; Kou, C; Gao, YF
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European Journal of Pharmaceutics and Biopharmaceutics
Exendin-4, a peptide analogue of glucagon-like peptide (GLP-1), has been developed for treatment of type 2 diabetes. Herein, the secretive exendin-4 peptide, expressed by methanol induction in Pichia pastoris, was purified to near homogeneity by Ni-NTA agarose chromatography. 103.6 mg of protein was obtained from 1 L of the supernatant and its purity was 96.1%. Subsequently, the PEGylated exendin-4 was prepared. The bioactivity of exendin-4 was determined by examining the glucose-lowering and insulin-releasing ability in plasma. Then, a safety evaluation was performed by histological examination of the main organs (liver, kidney and pancreas). PEGylated exendin-4 displayed glucose-lowering and insulin-stimulating action in vivo without obvious damage to the above organs. The results suggest that the P. pastoris expression could be used to produce large quantities of exendin-4 and PEGylation is a useful tool to maintain and enhance bioactivity of the peptide.
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