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Large-scale purification of human granulocyte-macrophage colony-stimulating factor expressed in Bombyx mori pupae

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Abstract

Human granulocyte-macrophage colony-stimulating factor (hGM-CSF) acts on many different kinds of cells, including monocytes, macrophages, granulocytes, eosinophils, and multipotential stem cells. To explore further explore pharmaceutical action, we expressed hGM-CSF by the Bombyx mori nucleopolyhedrovirus expression system in silkworm pupae. However, purifying recombinant proteins from silkworm pupae on a large scale has been a big challenge. To establish purification methods suitable for mass production, we tried two crude preparation methods: (NH4)2SO4 fractional precipitation and isoelectric precipitation with a combination of gel filtration andion-exchange chromatography. The isoelectric precipitation method was found to be more efficient. With this method, we eventually obtained approx 11.7 mg of 95% pure product from 1000 g of infected silkworm pupae. The recovery of purified protein was greatly increased, by approx 40%, compared with the other method. The biologic activity of this protein was determined up to 9.0×106 colony-forming units/mg in the final purified product.

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Author notes

  1. Yao-Zhou Zhang

    Present address: Institute of Biochemistry, Zhejiang Sci-Tech University, 310018, Hangzhou, China

Authors and Affiliations

  1. Institute of Biochemistry, Zhejiang Sci-Tech University, 310018, Hangzhou, China

    Jian Chen, Zuo-Ming Nie, Zheng-Bing Lü & Cheng-Zhi Xu

  2. College of Life Sciences, Zhejiang University, 310029, Hangzhou, China

    Cheng-Gang Zhu & Yong-Feng Jin

  3. Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, 200031, Shanghai, China

    Xiang-Fu Wu

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  1. Jian Chen
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Chen, J., Nie, ZM., Lü, ZB. et al. Large-scale purification of human granulocyte-macrophage colony-stimulating factor expressed in Bombyx mori pupae. Appl Biochem Biotechnol 141, 149–159 (2007). https://doi.org/10.1007/s12010-007-9217-1

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  • DOI: https://doi.org/10.1007/s12010-007-9217-1

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