[Preparation of novel magnetic dextran affinity adsorbents and their application to purify urokinase].
The reverse phase suspension and embedment technique were adopted to prepare magnetic dextran microsphere (MDMS). The dispersion medium was mixture of some organic solvents. Span-80 was used as stabilizer. The aqueous dextran with magnetic fluid was suspended in dispersion medium with epichlorohydrin as cross-linking reagent. The mixture was stirred for 30 minutes at room temperature and then heated at 70 degrees C for 4 hours, MDMS was thus obtained. MDMS was activated by epichlorohydrin on which 6-aminohexanoic acid, glycine or ethylene diamine was bonded as spacers. Then it was coupled with p-aminobenzamide, L-arginine methyl ester or guanidohexanoic acid and five magnetic affinity adsorbents were prepared. The MDMS was polydisperse particles with the size of 50-300 meshes and the content of Fe3O4 was about 6.2 per cent in the MDMS. Influence of some parameters such as viscosity and density of organic phase, the volume ratio of organic and aqueous phase, the quantity of surfactant and stirring speed on preparing MDMS was studied. Magnetic affinity adsorbents were used to purify crude urokinase in a bath mode and the effect of coupling reagents and ligands on results of purification was discussed. The bioactivity recovery was 40.0 to 60.7 per cent, the purification-fold was between 14.9 and 32.8, and the adsorptive capacity varies from 89 mg to 121 mg per milliliter of adsorbent.
Dong
Y S
YS
Department of Chemistry, Northwest University, Xi'an 710069, China.
Liang
F
F
Yu
X Y
XY
Guo
L A
LA
Chang
J H
JH
D000327:Adsorption; D003432:Cross-Linking Reagents; D003911:Dextrans; D008863:Microspheres; D013056:Spectrophotometry, Ultraviolet; D014568:Urokinase-Type Plasminogen Activator / Q000302:isolation & purification*
D003432:Cross-Linking Reagents; D003911:Dextrans; D014568:Urokinase-Type Plasminogen Activator