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US 12,618,058 B2Granted
Blends containing proteases
AssigneeRoche Diagnostics Operations, Inc.
InventorsWerner Hoelke, Michaela Fischer, Johann-Peter Thalhofer, Markus Weber
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Abstract
Described are compositions, in particular lyophilizates, containing proteolytic enzymes, and methods for producing the compositions. Typically these compositions contain one or more proteases with collagenase activity and a neutral protease, for example, thermolysin. The compositions are free of acetate salts. Surprisingly, such compositions can be dissolved in water more rapidly than lyophilized protease mixtures of the state of the art.
Claims16
A solid composition obtained or obtainable by the steps of
- (a) preparing a first homogeneous solution of an acetate-free preparation of a neutral protease (NP) in an aqueous acetate-free low-salt solution, the aggregate concentration of salt(s) in said low-salt solution being in the range of about 1 mM to about 250 mM;
- (b) adding a neutral salt to the first homogeneous solution of step (a) and dissolving the neutral salt, thereby making a stabilized solution, wherein said stabilized solution additionally comprises a buffer salt buffering in the range of about pH 6 to about pH 8.5, and wherein the stabilized solution further comprises calcium chloride;
- (c) mixing the stabilized solution of step (b) with an acetate-free preparation of one or more proteolytic enzymes with collagenase activity (C), and making a second homogeneous solution;
- and
- (d) freeze-drying the solution of step (c), thereby obtaining the solid composition, wherein:
- the neutral protease is thermolysin;
- the neutral salt is sodium chloride;
- and
- a total protein content in the second homogeneous solution is in the range of 1 mg/ml to 150 mg/ml, a concentration of the calcium chloride is in the range of 1 mM to about 10 mM, and a concentration of sodium chloride in the second homogeneous solution is in the range of about 50 mM to about 500 mM.
6 more elements
Dependencies are inferred from the claim text.