Cellulose as an inert matrix for presenting cytokines to target cells: production and properties of a stem cell factor-cellulose-binding domain fusion protein

• Published in: Biochemical journal Vol. 339; no. 2; pp. 429 - 434
• Main Authors: Doheny, J.G, Jervis, E.J, Guarna, M.M, Humphries, R.K, Warren, R.A.J, Kilburn, D.G
• Format: Journal Article
• Language: English
• Published: England 15-04-1999
• Subjects: Animals; binding; Cell Division; Cell Line; cellulose; Cellulose - metabolism; Cytokines - biosynthesis; Fluorescent Antibody Technique; growth factors; Humans; Mice; Microscopy, Confocal; Protein Binding; Recombinant Fusion Proteins - metabolism; Stem Cell Factor - biosynthesis; Stem Cell Factor - metabolism
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/0264-6021:3390429

A chimaera of stem cell factor (SCF) and a cellulose-binding domain from the xylanase Cex (CBDCex) effectively immobilizes SCF on a cellulose surface. The fusion protein retains both the cytokine properties of SCF and the cellulose-binding characteristics of CBDCex. When adsorbed on cellulose, SCF-CBDCex is up to 7-fold more potent than soluble SCF-CBDCex and than native SCF at stimulating the proliferation of factor-dependent cell lines. When cells are incubated with cellulose-bound SCF-CBDCex, activated receptors and SCF-CBDCex co-localize on the cellulose matrix. The strong binding of SCF-CBDCex to the cellulose surface permits the effective and localized stimulation of target cells; this is potentially significant for long-term perfusion culturing of factor-dependent cells. It also permits the direct analysis of the effects of surface-bound cytokines on target cells.