Compositions and methods of preparation thereof
View Patent ↗The present invention provides biological compositions and methods for making the same.
1. A method of preparing a fibrin composition, the method consisting of:
contacting a first portion of an anticoagulated whole blood, a fraction of the anticoagulated whole blood, or a component of the anticoagulated whole blood with a contact activation agent and an extrinsic coagulation pathway initiation agent to provide a coagulated mass in less than about 30 minutes;
extracting thrombin from the coagulated mass to provide a thrombin composition;
fractionating a second portion of the anticoagulated whole blood, the fraction of the anticoagulated whole blood, or the component of the anticoagulated whole blood to obtain a fibrinogen composition and a platelet plasma composition;
contacting the fibrinogen composition, platelet plasma composition, or a combination thereof with a recombinant protein composition;
contacting the thrombin composition with a stabilizing agent to provide a thrombin composition having a table-life of more than about 6 hours, wherein the anticoagulated whole blood, a fraction thereof or a component thereof is not contacted with the stabilizing agent during the generation of the coagulated mass;
contacting the thrombin composition, the fibrinogen composition and the platelet plasma composition to generate a fibrin composition; and
adding at least one micro-particle to the fibrin composition, wherein the at least one micro-particle includes at least one of the following: a pharmaceutical agent, a therapeutic agent, a medical agent, a biological agent, and any combination thereof;
adding at least one protecting group to protect one or more functional groups of at least one of the following: the thrombin composition, the fibrinogen composition, the platelet plasma composition, and the fibrin composition, wherein the protecting group includes at least one of the following: a carbonyl protecting group, an alcohol protecting group, and an amine protecting group.
2. The method of claim 1 , wherein the stabilizing agent comprises a polyol, PEG, ammonium sulfate, a non-polar solvent, a polar solvent, a methyl isobutyl ketone alcohol, glycol, tricloroacetic acid, acetate salt, or any combination thereof.
3. The method of claim 2 , wherein the stabilizing agent comprises ethanol.
4. The method of claim 3 , wherein about 8% to about 25% volume/volume ethanol is added to the thrombin composition.
5. The method of claim 4 , wherein about 10% volume/volume ethanol is added to the thrombin composition.
6. The method of claim 1 , wherein the thrombin composition has a table-life of more than 12 hours.
7. The method of claim 6 , wherein the thrombin composition has a table-life of from more than 12 up to 24 hours.
8. The method of claim 1 , wherein the coagulated mass is generated in less than about 10 minutes.
9. The method of claim 8 , wherein the coagulated mass is generated in less than about 5 minutes.
10. The method of claim 9 , wherein the coagulated mass is generated in less than about 3 minutes.
11. The method of claim 10 , wherein the coagulated mass is generated in about 1 minute to about 3 minutes.
12. The method of claim 1 , wherein the first portion is a portion of the anticoagulated whole blood, a portion of the fraction of the anticoagulated whole blood or a portion of the component of the anticoagulated whole blood and comprises platelet rich plasma (PRP), platelet poor plasma (PPP), or a combination thereof.
13. The method of claim 12 , wherein the first portion is a portion of the anticoagulated whole blood, a portion of the fraction of the anticoagulated whole blood or a portion of the component of the anticoagulated whole blood and comprises platelet rich plasma (PRP).
14. The method of claim 12 , wherein the first portion is a portion of the anticoagulated whole blood, a portion of the fraction of the anticoagulated whole blood or a portion of the component of the anticoagulated whole blood and comprises platelet poor plasma (PPP).
15. The method of claim 1 , wherein the second portion is a portion of the anticoagulated whole blood, a portion of the fraction of the anticoagulated whole blood or a portion of the component of the anticoagulated whole blood and comprises platelet rich plasma (PRP), platelet poor plasma (PPP), or a combination thereof.
16. The method of claim 15 , wherein the second portion is a portion of the anticoagulated whole blood, a portion of the fraction of the anticoagulated whole blood or a portion of the component of the anticoagulated whole blood and comprises platelet rich plasma (PRP).