| 摘要 |
PURPOSE:To obtain human granulocytes colony stimulation factor of high purity by solubilizing a protein having human granular colony stimulation factor activity, which is produced by the recombinant DNA technique under reductive conditions and oxidizing to form S-S bonds, removing pyrogenic substances and purifying. CONSTITUTION:A protein having an activity of human granulocytes colony stimulation factor (abbreviated to G-CSF) is produced using the recombinant DNA technique. The water-insoluble granules is solubilized by adding a modifier such as urea and a reducing agent such as 2-mercaptoethanol. Then, the modifier is removed, oxidized glutathione or cystine is added to effect oxidation to form a disulfide bond in the position corresponding to the disulfide bond in the natural G-CSF. Then, pyrogenic substances are removed from the mixture by silica gel column chromatography. Then, the protein having the activity of unmodified type of oxidized monomer G-CSF is isolated and purified by ultra-filtration, anion-exchange chromatography or the like. |
