Production of mitogen-contamination free alginates with variable ratios of mannuronic acid to guluronic acid by free flow electrophoresis.
Commercial alginates consisting of variable homopolymeric regions of beta-D-mannuronic acid and alpha-L-guluronic acid, interspaced with regions of alternating blocks, are potent stimulators of macrophages and lymphocytes. Therefore, inflammatory reactions and fibrotic overgrowth of the beads result if Langerhans islets are encapsulated in raw alginate hydrogel beads (cross-linked with divalent cations). The result is random failure of the islets some time after transplantation. Analysis of raw alginates by using free flow electrophoresis demonstrated that commercial alginates contained at least 10-20 fractions (characterized by different electrophoretic mobilities) which showed mitogenic activity. These fractions could be quantitatively separated from the alginic acids by free flow electrophoresis on a preparative scale. The purified alginates cross-linked with Ca2+ ions exhibited no mitogenic reactions as proved by an in vitro assay. In addition, examination of purified Ba2+ alginate beads implanted intraperitoneally in rats or mice for three weeks showed no fibrotic overgrowth in contrast to implants made from unpurified alginate.
Zimmermann
U
U
Institut für Biotechnologie, Universität Würzburg, Germany.
Klöck
G
G
Federlin
K
K
Hannig
K
K
Kowalski
M
M
Bretzel
R G
RG
Horcher
A
A
Entenmann
H
H
Sieber
U
U
Zekorn
T
T
D000464:Alginates / Q000008:administration & dosage / Q000032:analysis / Q000302:isolation & purification*; D000818:Animals; D003956:Dialysis; D004586:Electrophoresis / Q000379:methods; D005355:Fibrosis; D006603:Hexuronic Acids / Q000032:analysis*; D007249:Inflammation / Q000139:chemically induced; D007274:Injections, Intraperitoneal; D051379:Mice; D008863:Microspheres; D008934:Mitogens / Q000032:analysis*; D051381:Rats; D014574:Uronic Acids / Q000032:analysis*
D000464:Alginates; D006603:Hexuronic Acids; D008934:Mitogens; D014574:Uronic Acids; C007896:guluronic acid; C008324:mannuronic acid