Influence of blood proteins on biomedical analysis. VIII. Attempts at purification of gliclazide-degrading factor in human serum.
Gliclazide(hypoglycemic drug having sulfonylurea structure)-degrading activity was found in fraction M(macroglobulin, Fr. M) obtained from pooled human serum by gel filtration using a Sephadex G-150 column. The main degrading activity was in the fraction eluted from the Fr. M-subjected DEAE-cellulose column with 0.4 M phosphate buffer (pH 5.2), and the gliclazide-degrading protein localized around alpha 2 to beta-globulin on an electrophoretic pattern using a cellulose acetate membrane. The degrading activity was enhanced about two-fold by lyophilizing Fr. M solution containing a higher sodium phosphate (Na2HPO4-NaH2PO4), over 0.27 M. This indicates that the appearance and enhancement of the degrading activity required the combination of the lyophilization of the sample solution and a certain initial concentration of sodium phosphate prior to lyophilization.
K
K
Kobayashi
A
A
Hase
M
M
Kimura
T
T
Sakoguchi
M
M
Shimosawa
A
A
Matsuoka
D000510:Alpha-Globulins / Q000302:isolation & purification; D001609:Beta-Globulins / Q000302:isolation & purification; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004588:Electrophoresis, Cellulose Acetate; D005612:Freeze Drying; D005907:Gliclazide / Q000097:blood* / Q000378:metabolism; D006801:Humans; D010710:Phosphates / Q000494:pharmacology; D011487:Protein Conformation; D013453:Sulfonylurea Compounds / Q000097:blood*; D014462:Ultrafiltration
D000510:Alpha-Globulins; D001609:Beta-Globulins; D010710:Phosphates; D013453:Sulfonylurea Compounds; D005907:Gliclazide; C018279:sodium phosphate