Efficiency of a smooth desorption procedure for the purification of barley beta-amylase using immunoaffinity chromatography.
Immunoaffinity chromatography was used for a one step purification procedure of beta-amylase from the G25 Sephadex gel filtrated fraction of whole barley protein extracts. The immunoglobulin G (IgG) fraction of an anti-barley beta-amylase immune serum was immobilized on Ultrogel. A gentle desorption procedure was used, combining distilled water elution with an interrupted elution. The quality of the purification was assayed by using cross immunoelectrophoresis with a polyspecific anti-barley protein immune serum. The extent of the damaging effect of this procedure was evaluated on the specific activity of the enzyme and on its polymorphism, as displayed by isoelectric focusing. The results underline the efficiency of the purification procedure and its low denaturing effect on the beta-amylase. This opens new possibilities for some aspects of the enzyme study and for the purification of other biologically active proteins.
D000681:Amylases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002523:Edible Grain / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D001467:Hordeum / Q000201:enzymology*; D007120:Immunochemistry; D010940:Plant Proteins / Q000032:analysis; D001614:beta-Amylase / Q000302:isolation & purification*
D010940:Plant Proteins; D000681:Amylases; D001614:beta-Amylase