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Development of efficient method for purified recombinant bovine granulocyte-macrophage colony-stimulating factor production with baculovirus-silkworm gene expression system

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Abstract

Recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF) was produced by the baculovirus-silkworm expression system. It was purified to 98% by (NH4)2SO4, followed by a three-step column chromatography with silica gel, ion exchange resin and a metal chelate column. The specific activity of purified rboGM-CSF was 1.6–6.3 × 106 ED50 mg−1. By this method, the specific activity was raised 160-fold and 21% of the expressed rboGM-CSF was recovered.

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Acknowledgments

This work was supported by two Grants: the Recombinant Cytokine Project (RCP2002-1310) and the Insect Factory Project (No. 3106), provided by the Ministry of Agriculture, Forestry, and Fisheries of Japan.

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Authors and Affiliations

  1. Katakura Industries Co., Ltd., 1548 Shimo-okutomi, Sayama, Saitama, 350-1332, Japan

    Hidekazu Nagaya, Toshimichi Kanaya & Yoneko Tobita

  2. Kitayama Labes Co., Ltd., 6556-1 Nishiharuchika, Ina, Nagano, 399-4431, Japan

    Toshimichi Kanaya

  3. National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan

    Yuichi Yokomizo & Shigeki Inumaru

  4. Department of Molecular Immunology, School of Agriculture and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Takashi Onodera

Authors
  1. Hidekazu Nagaya
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  2. Toshimichi Kanaya
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  3. Yoneko Tobita
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  4. Yuichi Yokomizo
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  5. Shigeki Inumaru
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  6. Takashi Onodera
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Correspondence to Hidekazu Nagaya.

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Nagaya, H., Kanaya, T., Tobita, Y. et al. Development of efficient method for purified recombinant bovine granulocyte-macrophage colony-stimulating factor production with baculovirus-silkworm gene expression system. Biotechnol Lett 30, 41–45 (2008). https://doi.org/10.1007/s10529-007-9506-3

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  • DOI: https://doi.org/10.1007/s10529-007-9506-3

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