Thrombin isolated from blood and blood fractions

1. A method of preparing a solution comprising thrombin, the method comprising:

precipitating fibrinogen from a liquid comprising platelet-poor plasma or whole blood;

removing precipitated fibrinogen from the liquid by centrifugation or filtration to form a post-precipitation liquid;

incubating the post-precipitation liquid with calcium and a plurality of beads to activate a portion of the post-precipitation liquid to generate a solution comprising thrombin; and

separating the solution comprising thrombin from the beads.

2. The method of claim 1 , wherein the liquid comprises platelet-poor plasma.

3. The method of claim 1 , wherein the liquid comprising platelet-poor plasma or whole blood further comprises an anticoagulant.

4. The method of claim 3 , wherein the anticoagulant comprises acid-citrate-dextrose solution.

5. The method of claim 1 , wherein the precipitating comprises adding an organic solvent to the liquid comprising platelet-poor plasma or whole blood.

6. The method of claim 5 , wherein the organic solvent is methanol, ethanol, acetone, or combinations thereof.

7. The method of claim 1 , wherein the incubating comprises incubating for about 5 to about 10 minutes.

8. The method of claim 1 , wherein the liquid comprising platelet-poor plasma or whole blood further comprises a chelator and the calcium is present in an amount greater than an amount that can be complexed by the chelator.

9. The method of claim 1 , wherein the beads are glass beads, polystyrene beads, or combinations thereof.

10. The method of claim 1 , wherein separating the solution comprising thrombin from the beads comprises centrifuging the beads and post-precipitation liquid and removing a portion of the post-precipitation liquid supernatant as the solution comprising thrombin.

11. The method of claim 1 , further comprising at least one of: adding glycerin, a polyol, or alcohol to the solution comprising thrombin; adding a chelator to complex calcium in the solution comprising thrombin; adjusting the pH of the solution comprising thrombin to below 7; and reducing the temperature of the solution comprising thrombin.

12. The method of claim 1 , wherein:

precipitating fibrinogen from a liquid comprising platelet-poor plasma or whole blood comprises adding ethanol to platelet-poor plasma;

removing precipitated fibrinogen from the liquid to form a post-precipitation liquid comprises filtering the liquid;

incubating the post-precipitation liquid with calcium and a plurality of beads to activate a portion of the post-precipitation liquid to generate a solution comprising thrombin comprises mixing the post-precipitation liquid, calcium chloride, and beads; and

separating the solution comprising thrombin from the beads comprises centrifuging the beads and post-precipitation liquid and removing a portion of the post-precipitation liquid supernatant as the solution comprising thrombin.

13. The method of claim 1 , wherein

precipitating fibrinogen from a liquid comprising platelet-poor plasma or whole blood comprises drawing platelet-poor plasma or whole blood into a syringe containing ethanol, wherein the syringe further contains an anticoagulant when the liquid comprises whole blood;

removing precipitated fibrinogen from the liquid to form a post-precipitation liquid comprises coupling a filter to the syringe and filtering the liquid into a tube comprising the calcium and the plurality of beads;

incubating the post-precipitation liquid with calcium and a plurality of beads to activate a portion of the post-precipitation liquid to generate a solution comprising thrombin comprises mixing the post-precipitation liquid, calcium chloride, and beads in the tube; and

separating the solution comprising thrombin from the beads comprises centrifuging the beads and post-precipitation liquid and removing a portion of the post-precipitation liquid supernatant as the solution comprising thrombin.

14. The method of claim 13 , further comprising concentrating the solution comprising thrombin.

15. The method of claim 14 , wherein the beads are polyacrylamide beads, and wherein concentrating the solution comprising thrombin comprises incubating the solution with polyacrylamide beads.

16. The method of claim 1 , further comprising:

applying the solution comprising thrombin to a site on a subject.

17. The method of claim 16 , wherein the solution comprising thrombin is prepared from platelet-rich plasma or whole blood obtained from the subject.

18. A method of preparing a solution comprising thrombin from a whole blood sample in a container having a mixing assembly, the method comprising:

collecting a whole blood sample;

precipitating fibrinogen in the whole blood sample;

removing precipitated fibrinogen from the whole blood sample to form a post-precipitation whole blood sample, by filtering the whole blood sample or centrifuging the whole blood sample to sediment precipitated fibrinogen;

introducing the post-precipitation whole blood sample into the container;

mixing the post-precipitation whole blood sample with a calcium salt and with activating beads; and

withdrawing a volume of a solution comprising thrombin from the container after mixing the post-precipitation whole blood sample with the calcium salt and activating beads.

19. The method of claim 18 , wherein the precipitating comprises adding an organic solvent selected from the group consisting of methanol, ethanol, acetone, and combinations thereof.

20. The method of claim 18 , further comprising admixing ethanol to the mixed post-precipitation whole blood sample to achieve a concentration of about 4 percent (vol.) to about 7 percent (vol.) ethanol in the mixed post-precipitation whole blood sample.

21. The method of claim 18 , wherein the post-precipitation whole blood sample is mixed with polyacrylamide beads.

22. The method of claim 18 , further comprising:

applying the solution comprising thrombin to a site on a subject.

23. The method of claim 22 , wherein the solution comprising thrombin is prepared from a whole blood sample collected from the subject.