Cellulose as an inert matrix for presenting cytokines to target cells: production and properties of a stem cell factor-cellulose-binding domain fusion protein.
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.
Doheny
J G
JG
Department of Microbiology and Immunology, Biotechnology Laboratory, The University of British Columbia, 300-6174 University Boulevard, Vancouver, British Columbia, Canada V6T 1Z3.
Jervis
E J
EJ
Guarna
M M
MM
Humphries
R K
RK
Warren
R A
RA
Kilburn
D G
DG
D000818:Animals; D002455:Cell Division; D002460:Cell Line; D002482:Cellulose / Q000378:metabolism*; D016207:Cytokines / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D019089:Stem Cell Factor / Q000096:biosynthesis* / Q000378:metabolism
D016207:Cytokines; D011993:Recombinant Fusion Proteins; D019089:Stem Cell Factor; D002482:Cellulose