Abstract
Objective
To produce a therapeutic protein (endostatin) by fusion with two fragments of the carboxyl-terminal peptide (CTP) of the human chorionic gonadotropin β-subunit in Pichia pastoris.
Results
Two CTP sequences were fused to the C-terminal of human endostatin, and the fusion protein (endo-CTP) was expressed by P. pastoris. Endo-CTP inhibited proliferation of endothelial cells with an IC50 of 7 μg ml−1, and 30 % of cells were annexin V-positive after treatment with 20 μg endo-CTP ml−1 for 48 h. Migration of endothelial cells was inhibited by endo-CTP in a concentration-dependent manner. The half-life of endo-CTP in Sprague–Dawley rats was much longer than that of its commercial counterpart (Endostar).
Conclusion
A long-acting endostatin can be produced using CTP technology.
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Acknowledgments
This study was supported by National Nature Science Foundation (81572979), National Key Basic Research Program (2015CB931800), Shanghai Committee of Science and Technology (12431900202) and School of Pharmacy, Fudan University & The Open Project Program of Key Lab of Smart Drug Delivery (Fudan University), Ministry of Education, China (SDD2013-02).
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Baolong Wang and Xin Wang contributed equally to the work.
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Wang, B., Wang, X., Wayne, C. et al. Production of a therapeutic protein by fusing it with two fragments of the carboxyl-terminal peptide of human chorionic gonadotropin β-subunit in Pichia pastoris . Biotechnol Lett 38, 801–807 (2016). https://doi.org/10.1007/s10529-016-2038-y
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DOI: https://doi.org/10.1007/s10529-016-2038-y