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US 4,729,957 AGranted
Process for manufacture of L-asparaginase from erwinia chrysanthemi
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorsShwu-Maan Lee, John T. Ross, Marie H. Wroble
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Abstract
A process for the recovery and purification of L-asparaginase from Erwinia chrysanthemi is disclosed. The process involves the preparation of cellular acetone powder extract followed by either an ion exchange and affinity chromatography purification steps or by affinity chromatography alone. The column eluent is then dialyzed to produce substantially pure L-asparaginase.
Claims7
A process for purifying L-asparaginase from Erwina comprising the steps of:
- (1). Harvesting cells from both;
- (2). Permealizing the Erwina by mixing with acetone and passing acetone/cell
- mixture through a filter press at least once;
- (3). Rinsing the acetone/cell cake with buffer in the neutral pH range
- (pH 6.5-7.5) to remove acetone;
- (4). Rinsing cells with buffer in alkaline range;
- (5). Pass cells from step 4 through a cell press to extract the enzyme;
- (6). Clarify the enzyme containing fluid obtained in step 5 by in-line passage
- through filter bag loaded with Cell Debris Remover;
- (7). Purifying the enzyme on an affinity chromatography column.
7 more elements
Dependencies are inferred from the claim text.