Development of efficient method for purified recombinant bovine granulocyte-macrophage colony-stimulating factor production with baculovirus-silkworm gene expression system.
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.
Nagaya
Hidekazu
H
Katakura Industries Co., Ltd., 1548 Shimo-okutomi, Sayama, Saitama 350-1332, Japan. h.nagaya@katakura.co.jp
Kanaya
Toshimichi
T
Tobita
Yoneko
Y
Yokomizo
Yuichi
Y
Inumaru
Shigeki
S
Onodera
Takashi
T
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D002417:Cattle / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor