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A strategy for fusion expression and preparation of functional glucagon-like peptide-1 (GLP-1) analogue by introducing an enterokinase cleavage site

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Abstract

KGLP-1, a 31-amino acid glucagon-like peptide-1 (GLP-1) analogue, has a great therapeutic potential for anti-diabetes. In this work, a strategy for expression and purification of functional KGLP-1 peptide has been established. KGLP-1 cDNA was fused with glutathione S-transferase (GST), with an enterokinase cleavage site in the fusion junction. The recombinant fusion protein GST–KGLP-1 was affinity purified via the GST-tag, and then digested with enterokinase. The resulting GST part as well as the enzymes were eliminated by ultra-filtration followed by size exclusion chromatograph. The yield of purified KGLP-1 was approximately 12.1 mg/L, with purity of 96.18 %. The recombinant KGLP-1 was shown to have similar bioactivity as native GLP-1 when evaluated in a Chinese hamster ovary cell line expressing a GLP-1 receptor-egfp reporter gene.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program 2012AA021505) and the National Natural Science Foundation of China (21002052).

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Authors and Affiliations

  1. College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, 94 Weijin Road, Tianjin, 300071, People’s Republic of China

    Yang Liu, Limei Ren, Lingmiao Ge, Qingxin Cui, Xiaofang Cao, Yuanyuan Hou, Fang Bai & Gang Bai

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  1. Yang Liu
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  2. Limei Ren
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  3. Lingmiao Ge
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  4. Qingxin Cui
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  5. Xiaofang Cao
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  6. Yuanyuan Hou
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  7. Fang Bai
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  8. Gang Bai
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Corresponding author

Correspondence to Fang Bai.

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Liu, Y., Ren, L., Ge, L. et al. A strategy for fusion expression and preparation of functional glucagon-like peptide-1 (GLP-1) analogue by introducing an enterokinase cleavage site. Biotechnol Lett 36, 1675–1680 (2014). https://doi.org/10.1007/s10529-014-1526-1

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  • DOI: https://doi.org/10.1007/s10529-014-1526-1

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