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Method for purifying porcine mature interleukin-18 from silkworm haemolymph

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Abstract

Recombinant porcine mature interleukin-18 (rPomIL-18) has been purified from the haemolymph of silkworms. After co-infection of two recombinant baculoviruses (BmAcpVL1392-IL-18-His and BmAcpVL1392-casp-1) into the silkworm, rPomIL-18 was produced and secreted into the haemolymph. Polyethylene glycol (PEG) 6000 was added to the haemolymph at 8% (v/w) to precipitate storage proteins which would otherwise bind non-specifically to the metal chelating column and the supernatant then was applied to Sepharose bonded with Ni2+. rPomIL-18 was eluted from the column using 100 mm imidazole buffer at pH 8 with a purity of 93.6%. Approximately 5.3 mg purified rPomIL-18 was obtained from 22 ml haemolymph. It could induce interferon-γ formation from porcine peripheral blood mononuclear cells.

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Authors and Affiliations

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

    Hidekazu Nagaya, Chiaki Enomoto & Sayaka Matsumoto

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

    Yoshihiro Muneta, Yuichi Yokomizo & Yasuyuki Mori

Authors
  1. Hidekazu Nagaya
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  2. Yoshihiro Muneta
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  3. Chiaki Enomoto
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  4. Sayaka Matsumoto
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  5. Yuichi Yokomizo
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  6. Yasuyuki Mori
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Nagaya, H., Muneta, Y., Enomoto, C. et al. Method for purifying porcine mature interleukin-18 from silkworm haemolymph. Biotechnology Letters 26, 869–873 (2004). https://doi.org/10.1023/B:bile.0000025894.87314.2d

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  • DOI: https://doi.org/10.1023/B:bile.0000025894.87314.2d

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