| Abstrakt: |
Human granulocyte-macrophage colony-stimulating factor (hGM-CSF)acts on many different kinds of cells, including monocytes, macrophages,granulocytes, eosinophils, and multipotential stem cells. To explore furtherexplore pharmaceutical action, we expressed hGM-CSF by the Bombyx morinucleopolyhedrovirus expression system in silkworm pupae. However,purifying recombinant proteins from silkworm pupae on a large scale hasbeen a big challenge. To establish purification methods suitable for massproduction, we tried two crude preparation methods: (NH4)2SO4 fractionalprecipitation and isoelectric precipitation with a combination of gel filtrationand ion-exchange chromatography. The isoelectric precipitation method wasfound to be more efficient. With this method, we eventually obtained approx11.7 mg of 95% pure product from 1000 g of infected silkworm pupae. Therecovery of purified protein was greatly increased, by approx 40%, comparedwith the other method. The biologic activity of this protein was determinedup to 9.0 × 106 colony-forming units/mg in the final purified product. [ABSTRACT FROM AUTHOR] |
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