Affinity separation of human plasma gelsolin on Affi-Gel Blue.
Human plasma gelsolin was specifically eluted with 1 mM adenosine 5'-triphosphate from an Affi-Gel Blue column. Since the ionic strength of sodium chloride required to elute the protein from the dye column was much higher than that of 1 mM adenosine 5'-triphosphate, the binding of plasma gelsolin with the dye-ligand appeared to be biospecific. Taking advantage of this affinity interaction, we have developed a revised purification method of human plasma gelsolin. The purification included ammonium sulfate precipitation, diethylaminoethyl-Sepharose chromatography, Affi-Gel Blue chromatography, and Phenyl-Sepharose chromatography. The method allowed a reproducible purification of the protein to apparent homogeneity, producing a 331-fold purification with a yield of 6%.
Yamamoto
H
H
Third Department of Internal Medicine, Osaka University School of Medicine.
Terabayashi
M
M
Egawa
T
T
Hayashi
E
E
Nakamura
H
H
Kishimoto
S
S
D000199:Actins / Q000302:isolation & purification; D000645:Ammonium Sulfate; D002135:Calcium-Binding Proteins / Q000097:blood* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D018260:Gelsolin; D006801:Humans; D015151:Immunoblotting; D008840:Microfilament Proteins / Q000097:blood* / Q000302:isolation & purification; D014227:Triazines
D000199:Actins; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; D014227:Triazines; C012842:Cibacron Blue F 3GA; D000645:Ammonium Sulfate