Materials and methods for producing blood products
Provided herein are materials and methods for the preparation of blood products. In one aspect, provided herein is a composition including platelets or platelet derivatives and an aqueous medium, wherein the aqueous medium has a protein concentration less than 50% of the protein concentration of donor apheresis plasma.
1. A process for preparing a thermally-treated platelet composition, the process comprising:
performing tangential flow filtration (TFF) of a platelet composition with a preparation agent comprising a buffering agent, trehalose in an amount in the range of 10 mM to 500 mM, and polysucrose in an amount in the range of 3% to 7%, thereby preparing a TFF-treated composition comprising at least 1000×10 3 platelets/μl in an aqueous medium having less than or equal to 7.5% plasma protein and having less than 5.0% microparticles by scattering intensity,
freeze drying the TFF-treated composition comprising platelets in the aqueous medium to form a freeze-dried platelet composition; and
heating the freeze-dried platelet composition at a temperature in the range of 60° C. to 85° C. for at least 1 hour to not more than 36 hours to thermally treat the freeze-dried platelets in the freeze-dried platelet composition to form the thermally-treated platelet composition, wherein thermally-treated platelets in the thermally-treated platelet composition have a potency of at least 1.5 thrombin generation potency units (TGPU) per 10 6 thermally-treated platelets.
2. The process of claim 1 , wherein the platelet composition has a plasma protein concentration of about 60 to about 80 mg/mL and the thermally-treated platelet composition has a plasma protein concentration of less than 6 mg/ml.
3. The process of claim 1 , wherein the buffering agent comprising comprises HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid.
4. The process of claim 1 , wherein the preparation agent comprises an organic solvent comprising ethanol in the range of 0.1% to 5.0%.
5. The process of claim 1 , wherein at least 55% of the thermally-treated platelets in the thermally-treated platelet composition are CD41 positive.
6. The process of claim 1 , wherein at least 70% of the thermally-treated platelets in the thermally-treated platelet composition are annexin V positive.
7. The process of claim 1 , wherein at least 8% of the thermally-treated platelets in the thermally-treated platelet composition are CD47 positive.
8. The process of claim 1 , wherein at least 80% of the thermally-treated platelets in the thermally-treated platelet composition are CD62 positive.
9. The process of claim 1 , wherein at least some of the thermally-treated platelets in the thermally-treated platelet composition have fibrinogen associated with their cell membrane.
10. The process of claim 1 , wherein the thermally-treated platelets in the thermally-treated platelet composition, when at a concentration of about 4.8×10 3 particles/μL generate a thrombin peak height (TPH) of at least 25 nM when in the presence of a reagent containing tissue factor and phospholipids.
11. The process of claim 1 , wherein the thermally-treated platelets in the thermally-treated platelet composition, when at a concentration of at least about 70×10 3 particles/μL, produce an occlusion time of less than 14 minutes in a total thrombus-formation analysis system (T-TAS) assay.
12. The process of claim 1 , wherein the thermally-treated platelets in the thermally-treated platelet composition have a shelf-life of 2-3 years.
13. The process of claim 1 , wherein the thermally-treated platelet composition is:
a) negative for HLA Class I antibodies based on a regulatory agency approved test for HLA Class I antibodies;
b) negative for HLA Class II antibodies based on a regulatory agency approved test for HLA Class II antibodies;
c) negative for HNA antibodies based on a regulatory agency approved test for HNA antibodies; or
d) two or more of a), b), and c).
14. The process of claim 1 , wherein the aqueous medium has a plasma protein concentration less than or equal to 50% of the plasma protein concentration of donor apheresis plasma that was the source of the platelet composition.
15. The process of claim 1 , wherein the preparation agent has a pH of 6.0 to 7.4.
16. The process of claim 1 , wherein the process does not comprise treatment of the platelet composition with an agent capable of altering nucleic acids of a pathogen prior to performing the TFF.
17. The process of claim 1 , further comprising a pathogen reduction step before the step of performing TFF.
18. The process of claim 1 , wherein trehalose in the preparation agent is in an amount in the range of 50 mM to 500 mM and the polysucrose is in an amount in the range of 3% to 6%.
19. The process of claim 1 , wherein the TFF-treated composition comprises microparticles in an amount below 3.0% by scattering intensity and wherein at least 80% of the thermally-treated platelets in the thermally-treated platelet composition are CD42 positive.
20. The process of claim 1 , wherein the polysucrose in the preparation agent is in an amount in the range of 3% to 6%.
21. The process of claim 1 , wherein the TFF-treated composition comprises platelets in the range of 1000×10 3 to 4000×10 3 platelets/μl in the aqueous medium.
22. The process of claim 1 , wherein the TFF-treated composition comprises at least 2000×10 3 platelets/μl in the aqueous medium.
23. The process of claim 1 , wherein the heating of the freeze-dried platelet composition is done at a temperature in the range of 70° C. to 85° C.
24. The process of claim 1 , wherein the thermally-treated platelet composition is:
a) negative for HLA Class I antibodies based on a regulatory agency approved test for HLA Class I antibodies;
b) negative for HLA Class II antibodies based on a regulatory agency approved test for HLA Class II antibodies; and
c) negative for HNA antibodies based on a regulatory agency approved test for HNA antibodies.
25. The process of claim 1 , wherein the TFF is performed using a membrane with a pore size in the range of 0.45 to 0.65μ, wherein the TFF-treated composition has less than 3.0% microparticles by scattering intensity, and wherein the heating the freeze-dried platelet composition is performed at a temperature in the range of 70° C. to 85° C. for between 6 hours and 36 hours to thermally treat the freeze-dried platelets to form the thermally-treated platelet composition.
26. The process of claim 25 , wherein the TFF is performed using a membrane with a pore size of 0.45μ.
27. The process of claim 1 , wherein the heating the freeze-dried platelet composition is performed at a temperature in the range of 75° C. to 85° C. for between 6 hours and 36 hours to thermally treat the freeze-dried platelets to form the thermally-treated platelet composition.
28. The process of claim 1 , wherein the step of performing TFF comprises diluting the platelet composition with a preparation agent to form a diluted platelet composition;
concentrating the diluted platelet composition to form a concentrated platelet composition such that the platelets in the concentrated platelet composition have a concentration in the range of 2000×10 3 cells/μl to 2500×10 3 cells/μl; and
performing TFF of the concentrated platelet composition with at least 2 dia-volumes (DV) of the preparation agent, thereby preparing the TFF-treated composition.
29. The process of claim 25 , wherein at least 80% of the thermally-treated platelets in the thermally-treated platelet composition are CD42 positive.