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利用杆状病毒-家蚕基因表达系统开发高效生产纯化重组牛粒细胞-巨噬细胞集落刺激因子的方法。

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

作者信息

Nagaya Hidekazu, Kanaya Toshimichi, Tobita Yoneko, Yokomizo Yuichi, Inumaru Shigeki, Onodera Takashi

机构信息

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

出版信息

Biotechnol Lett. 2008 Jan;30(1):41-5. doi: 10.1007/s10529-007-9506-3. Epub 2007 Aug 14.

DOI:10.1007/s10529-007-9506-3
PMID:17700995
Abstract

Recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF) was produced by the baculovirus-silkworm expression system. It was purified to 98% by (NH(4))(2)SO(4), 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 x 10(6) ED(50) mg(-1). By this method, the specific activity was raised 160-fold and 21% of the expressed rboGM-CSF was recovered.

摘要

重组牛粒细胞-巨噬细胞集落刺激因子（rboGM-CSF）由杆状病毒-家蚕表达系统产生。通过硫酸铵将其纯化至98%，随后依次用硅胶、离子交换树脂和金属螯合柱进行三步柱层析。纯化后的rboGM-CSF的比活性为1.6 - 6.3×10⁶ ED₅₀ mg⁻¹。通过该方法，比活性提高了160倍，并且回收了21%表达的rboGM-CSF。

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