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US 8,617,863 B2Granted
Composition, method, and kit for preparing plasmin
AssigneeGrifols Therapeutics Inc.
InventorsEdward Koepf, Thomas Zimmerman
Life of this patent
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Abstract
A streptokinase immobilized on a surface, in particular an immobilized plasmin-resistant streptokinase, and compositions, methods and kits of utilizing same for preparing plasmin are provided.
Claims20
A method for preparing plasmin, the method comprising:
- (a) contacting a composition comprising a plasminogen with a streptokinase immobilized on a matrix thereby converting the plasminogen to a plasmin,
- wherein the streptokinase is a streptokinase mutant characterized as capable of activating plasminogen to plasmin, yet resistant to plasmin degradation relative to its corresponding wild-type streptokinase;
- and
- wherein the streptokinase comprises an amino acid sequence having an amino acid residue other than lysine at a position corresponding to position 85, 412, or both positions, as shown in SEQ ID NO:1;
- and
- (b) purifying the plasmin by a method comprising contacting the composition with a plasmin-binding matrix so that the plasmin is retained by the plasmin-binding matrix, the plasmin-binding matrix having a molecule disposed thereon having affinity for the plasmin.
3 more elements
A method for preparing plasmin, the method comprising:
- (a) contacting a composition comprising a plasminogen with a streptokinase immobilized on a matrix thereby converting the plasminogen to a plasmin,
- wherein the streptokinase is a streptokinase mutant characterized as capable of activating plasminogen to plasmin, yet resistant to plasmin degradation relative to its corresponding wild-type streptokinase;
- wherein the streptokinase comprises an amino acid sequence having an amino acid residue other than lysine at a position corresponding to position 85, 412, or both positions, as shown in SEQ ID NO:1;
- and
- wherein the streptokinase comprises a polyhistidine affinity tag;
- and
- (b) purifying the plasmin by a method comprising contacting the composition with a plasmin-binding matrix so that the plasmin is retained by the plasmin-binding matrix;
- wherein the plasma-binding matrix is benzamidine-coupled to crosslinked beaded agarose.
5 more elements
Dependencies are inferred from the claim text.