Preparative purification of pig red blood cell hexokinase type III using a new efficient chromatographic support.
In this paper we report the purification of pig erythrocyte hexokinase type III, at preparative level, using 52 liters of starting material (hemolysate). This was possible using a new efficient anion exchanger support, the Toyopearl DEAE 650 M which allows completely to change the strategy of removing hemoglobin from hemolysates, permitting to handle large amounts of starting material and reducing work would have required months using conventional anion exchanger supports, to only 2-3 days. Furthermore, we have tested the binding of other red blood cell enzymes to the Toyopearl DEAE 650 M, showing a wider potential use of this chromatographic support for their purification at a preparative level.
Stocchi
V
V
Istituto di Chimica Biologica, Giorgio Fornaini, Università degli Studi, Urbino, Italy.
Masat
L
L
Biagiarelli
B
B
Piccoli
G
G
Palma
F
F
Cucchiarini
L
L
Dachà
M
M
D000818:Animals; D000837:Anion Exchange Resins; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004798:Enzymes / Q000032:analysis / Q000302:isolation & purification; D004912:Erythrocytes / Q000201:enzymology*; D006593:Hexokinase / Q000302:isolation & purification*; D007527:Isoenzymes / Q000302:isolation & purification*; D012117:Resins, Synthetic; D013552:Swine
D000837:Anion Exchange Resins; D004798:Enzymes; D007527:Isoenzymes; D012117:Resins, Synthetic; C075368:Toyopearl DEAE 650 M; D006593:Hexokinase