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Expression and purification of soluble recombinant Human Endostatin in Escherichia coli

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Abstract

Endostatin, a 20 kDa C-terminal fragment of collagen XVIII, is a specific inhibitor of endothelial cell proliferation and angiogenesis. In the present study, we produced soluble and biologically active recombinant human endostatin (rhEndostatin) in Escherichia coli by expressing via fusion with solubility-promoting peptides and optimizing the expression conditions. The rhEndostatin was expressed via fusion with glutathione S-transferase (GST) and NusA protein, respectively. It revealed that NusA protein enhanced the production of soluble rhEndostatin; but GST didn’t. By optimizing the expression conditions, the production of soluble NusA-rhEndostatin fusion protein was about 50% of total cellular proteins and about 90% of the products appeared in the cellular supernatant fraction. The soluble NusA-rhEndostatin fusion protein was purified by one-step hydrophobic interaction chromatography and NusA was removed by thrombin. Then rhEndostatin was purified by affinity chromatography and gel filtration chromatography. As a result, a simple and economical purification procedure for rhEndostatin isolation was obtained. The biological activity of the rhEndostatin was demonstrated in vitro using a human vascular endothelial cells (HuVECs) proliferation assay. Our study provides a feasible and convenient approach to produce soluble and biologically active rhEndostatin.

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China

    Cuihong Du, Xiaoping Yi & Yuanxing Zhang

  2. School of Biotechnology Engineering, Jimei University, Xiamen, 361021, China

    Cuihong Du

  3. Shandong Dongeejiao Group, Shandong, 252201, China

    Cuihong Du, Xiaoping Yi & Yuanxing Zhang

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  1. Cuihong Du
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  2. Xiaoping Yi
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  3. Yuanxing Zhang
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Correspondence to Cuihong Du.

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Du, C., Yi, X. & Zhang, Y. Expression and purification of soluble recombinant Human Endostatin in Escherichia coli . Biotechnol Bioproc E 15, 229–235 (2010). https://doi.org/10.1007/s12257-009-0100-5

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  • DOI: https://doi.org/10.1007/s12257-009-0100-5

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