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| Manufaction of a New High-glycosylated Fusion Protein NESP-Fc |
| YAN Li, WANG Kang, LI Rui-jian, BAI Yi, BAI Xian-hong, ZHOU Hai-ping |
| BioTech Pharmaceuticals Co., Ltd., Beijing 100176, China |
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Abstract New erythropoiesis stimulating protein (NESP), is a highly glycosylated rhEPO analogues containing five N-terminal sugar chain and twice as high in sialic acid residues than that of rhEPO. NESP has greater metabolic stability and three times the half-life of rhEPO. Fusion with Fc fragment can prolongs half-life of a protein in vivo. NESP was used to construct the new erythropoiesis stimulating protein-IgG2 Fc fragment fusion protein (NESP-Fc).After process optimization, the expression of NESP-Fc is about 1.4g/L. A series of studies confirmed that the fusion protein can form dimer, and the molecular weight of the dimer is about 130kDa. The NESP-Fc fusion protein can significantly promote the growth of UT-7 cells in vitro, and fusion with Fc didn't impact the biological function of NESP. In mouse model, NESP-Fc dose-dependently increased reticulocyte level. The half-life of NESP-Fc fusion protein can reach up to 56 hours in rats. These studies of NESP-Fc fusion protein proved that NESP-Fc could be potentially used as a medicine for anemia, and it also laid a solid foundation for future clinical trial and the final industrialization.
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Received: 26 November 2014
Published: 25 April 2015
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