Purification of human fibroblast interferon by ganglioside-sepharose 4B

Abstract

The cell surface is important in several phases of interferon action. When a cell is treated with an interferon to which it is sensitive, an antiviral state develops. During this process, interferon appears to bind to specific receptors on the cell surface. Studies over the past few years, indicate that receptors for interferon are composed of both glycoprotein and ganglioside components, and that key determinant within the receptor may be an oligosaccharide moiety. Using this interferon-ganglioside interaction, interferon was purified. At first, BSA was coupled to CNBr-activated sepharose 4B, thereafter, ganglioside was immibilized to BSA-sepharose 4B. With this ganglioside-BSA- sepharose 4B Beads, experiments were carried out. Human fibroblast interferon produced by human diploid fibroblast cell, was bound to this beads and released at the various conditions. Binding capacity of this beads was 4500 unit per ml of beads and amounts of bount interferon was maximized at the condition of 30$^\circ$C, 45 minutes incubation. Most effective conditions for releasing of bound interferon was 0.1M lactose and 0.02M N-acetylneuraminic acid in sodium phosphate buffer (0.02M, pH 7.4), and in this condition, recovery yield was 76.6\%.