High-performance purification of gelsolin from plasma using anion-exchange porous hollow-fiber membrane.
Gelsolin was purified from bovine plasma using an anion-exchange porous hollow-fiber membrane. The anion-change porous hollow-fiber membrane was prepared by radiation-induced graft polymerization of an epoxy-group-containing monomer, glycidyl methacrylate, and subsequent chemical modifications. Some of the epoxy groups of the polymer chain grafted onto the pore surface were converted into diethylamino groups, and the remaining epoxy groups were converted into 2-hydroxyethylamino groups. First, a gelsolin-containing dialyzed protein solution, prepared by pretreatments of ammonium sulfate precipitation and dialysis of plasma, was forced to permeate through the pores of an anion-exchange porous hollow-fiber membrane. Various proteins including gelsolin were adsorbed onto the anion-exchange polymer brush at a high rate with negligible diffusional mass-transfer resistance. Second, adsorbed gelsolin was specifically eluted by permeating 2mM calcium chloride. The amount of recovered gelsolin was 0.1 mg per 1 mL of plasma. Third, the remaining adsorbed proteins were quantitatively eluted with 1M sodium chloride, leading to a constant amount of recovered gelsolin during four cycles of purification. The total time required for gelsolin purification from 30 mL of bovine plasma was 11h, during which the time for selective adsorption of various proteins and affinity elution of gelsolin using the anion-exchange porous hollow-fiber membrane was 20 min.
Hagiwara
Kyohei
K
Department of Applied Chemistry and Biotechnology, Chiba University, 1-33 Yayoi, Inage-ku, Chiba 263-8522, Japan.
Yonedu
Shinji
S
Saito
Kyoichi
K
Shiraishi
Tomoyuki
T
Sugo
Takanobu
T
Tojyo
Tadashi
T
Katayama
Eisaku
E
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004852:Epoxy Compounds / Q000737:chemistry; D018260:Gelsolin / Q000097:blood* / Q000302:isolation & purification*; D008567:Membranes, Artificial; D008689:Methacrylates / Q000737:chemistry
D004852:Epoxy Compounds; D018260:Gelsolin; D008567:Membranes, Artificial; D008689:Methacrylates; C007870:glycidyl methacrylate