Method and formulation for stabilization of enzymes
AssigneeMolecular Biology Resources, Inc.
InventorsTherese A. De Rosier, Norberto B. de la Cruz, Richard K. Wilkosz
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Abstract
The present invention is directed to a method and a formulation for the stabilization of an enzyme for storage at temperatures above freezing. Specifically, the invention involves the use of formulated excipients that when added to an enzyme or mixture of enzymes and subsequently lyophilized renders stability. Upon reconstitution, the stabilized materials are useful in assays, diagnostic or molecular biology research kits, and other biological applications.
Claims66
A method for stabilizing one or more enzymes, or enzymatically active fragments thereof, for storage in the dry phase above freezing temperature, the method comprising:
- (i) combining said one or more enzymes or enzymatically active fragment thereof with an aqueous stabilizing solution comprising in combination a carrier protein, from about 3% to about 15% w/v of one or more disaccharides, from about 1% to about 10% w/v of one or more disaccharide derivatives, from about 0% to about 10% w/v of one or more sugar polymers, and a compatible buffer thereby producing a mixture;
- (ii) freezing the mixture obtained in step (i);
- and
- (iii) lyophilizing the frozen mixture obtained in step (ii) until substantially water-free, whereby a stabilized enzyme formulation is obtained.
A mixture for use in producing an aqueous formulation suitable for lyophilization, comprising:
- (i) a carrier protein;
- (ii) from about 3% to about 15% (w/v) of one or more disaccharides;
- (iii) from about 1% to about 10% (w/v) of one or more disaccharide derivatives;
- (iv) from about 0% to about 10% (w/v) of a sugar polymer;
- (v) a compatible buffer;
- and
- (vi) one or more enzymes.
A stabilized mixture of one or more enzymes, or enzymatically active fragments thereof, produced by a method comprising:
- (i) combining one or more enzymes, or enzymatically active fragments thereof, with an aqueous stabilizing solution comprising in combination a carrier protein, from about 1% to about 10% (w/v) of one or more disaccharide derivatives, from about 3% to about 15% (w/v) of one or more disaccharides, from about 0% to about 10% of one or more simple sugar polymers, and a compatible buffer, thereby producing a mixture;
- (ii) freezing said mixture;
- and
- (iii) lyophilizing said frozen mixture until substantially water-free, whereby a stabilized mixture of said one or more enzymes is obtained.
An aqueous enzyme formulation useful for stabilizing one or more enzymes, or enzymatically active fragments thereof, produced by a method comprising combining with an aqueous phase:
- (i) a carrier protein;
- (ii) from about 1% to about 10% (w/v) of one or more disaccharide derivatives;
- (iii) from about 3% to about 15% (w/v) of one or more disaccharides;
- (iv) from about 0% to about 10% of one or more simple sugar polymers;
- (v) a compatible buffer;
- and
- (vi) one or more enzymes, or enzymatically active fragments thereof.
A stabilized, substantially water-free mixture of one or more enzymes or enzymatically active fragments thereof comprising:
- (i) a carrier protein;
- (ii) from about 3% to about 15% (w/v) of one or more disaccharides;
- (iii) from about 1% to about 10% (w/v) of one or more disaccharide derivatives;
- (iv) from 0% to about 10% (w/v) of a sugar polymer;
- (v) a compatible buffer;
- and
- (vi) one or more enzymes.
Dependencies are inferred from the claim text.