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 comprising platelets in a preparation agent comprising a buffering agent, between 10 mM and 1000 mM of saccharides comprising a monosaccharide and/or a disaccharide, and polysucrose in an amount in the range of 3% to 7%, wherein the TFF is performed until a particular residual plasma protein endpoint is reached, thereby preparing a TFF-treated composition comprising at least 1000×10 3 platelets/μl in an aqueous medium having less than or equal to 15% 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 the thermally-treated platelets in the thermally-treated platelet composition are capable of generating thrombin in an in vitro thrombin formation assay.
2. The process of claim 1 , wherein at least 79% of the thermally-treated platelets in the thermally-treated platelet composition are CD42 positive.
3. The process of claim 1 , wherein the particular residual plasma protein endpoint is set at less than or equal to 7.5% residual plasma protein.
4. The process of claim 1 , wherein the particular residual plasma protein endpoint is set at less than or equal to 5% residual plasma protein.
5. The process of claim 1 , wherein an absorbance at 280 nm (A280) of the aqueous medium is measured to determine that the particular residual plasma protein endpoint is reached.
6. The process of claim 2 , wherein between 79% and 95% of the thermally-treated platelets in the thermally-treated platelet composition are CD42 positive, and at least 78% of the thermally-treated platelets in the thermally-treated platelet composition are CD41 positive.
7. The process of claim 2 , wherein at least 80% of the thermally-treated platelets in the thermally-treated platelet composition are CD42 positive, and at least 80% of the thermally-treated platelets in the thermally-treated platelet composition are CD41 positive.
8. The process of claim 2 , wherein the saccharides comprise the monosaccharide and the disaccharide, and wherein the disaccharide is selected from the group consisting of trehalose, sucrose, maltose, and combinations thereof, and the monosaccharide is selected from the group consisting of glucose, mannose, dextrose, xylose, and combinations thereof.
9. The process of claim 8 , wherein the disaccharide comprises trehalose in an amount in the range of 10 mM to 500 mM, and the monosaccharide comprises dextrose.
10. The process of claim 2 , wherein the saccharides comprise a disaccharide selected from the group consisting of trehalose, sucrose, maltose, and combinations thereof, and wherein the disaccharide is in an amount in the range of 10 mM to 500 mM.
11. The process of claim 10 , wherein the disaccharide is trehalose in an amount in the range of 50 mM to 500 mM.
12. The process of claim 2 , wherein the thermally-treated platelets in the thermally-treated platelet composition are capable of generating clots in an in vitro clotting assay, and promoting aggregation or co-aggregation in an in vitro aggregation or co-aggregation assay.
13. The process of claim 12 , 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.
14. The process of claim 2 , wherein the TFF is performed using a membrane with a pore size in the range of 0.45 to 0.65 μm, and wherein the TFF-treated composition has less than 3.0% microparticles by scattering intensity.
15. 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.
16. 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).
17. 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.
18. The process of claim 7 , wherein the TFF is performed using a membrane with a pore size in the range of 0.45 to 0.65 μm, and wherein the TFF-treated composition has less than 3.0% microparticles by scattering intensity, and wherein the thermally-treated platelets in the thermally-treated platelet composition are capable of generating clots in an in vitro clotting assay.
19. The process of claim 7 , wherein the thermally-treated platelets in the thermally-treated platelet composition:
a) 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;
b) 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; or
c) both a) and b).
20. The process of claim 15 , wherein the thermally-treated platelets in the thermally-treated platelet composition:
a) 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; and
b) 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.
21. The process of claim 15 , wherein the thermally-treated platelets in the thermally-treated platelet composition comprises:
a) at least 55% thermally-treated freeze-dried platelets positive for CD41;
b) at least 70% thermally-treated freeze-dried platelets positive for annexin V;
c) at least 8% thermally-treated freeze-dried platelets positive for CD47;
d) at least 80% thermally-treated freeze-dried platelets positive for CD62; or
e) two or more of a), b), c), and d).
22. The process of claim 21 , wherein the thermally-treated platelets in the thermally-treated platelet composition comprises:
b) at least 85% thermally-treated freeze-dried platelets positive for annexin V; and
d) at least 85% thermally-treated freeze-dried platelets positive for CD62 positive.
23. The process of claim 21 , wherein the thermally-treated platelets in the thermally-treated platelet composition comprises:
a) at least 75% thermally-treated freeze-dried platelets positive for CD41;
b) at least 85% thermally-treated freeze-dried platelets positive for annexin V; and
d) at least 85% thermally-treated freeze-dried platelets positive for CD62.
24. The process of claim 1 , wherein the buffering agent comprising comprises HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, and the preparation agent further comprises an organic solvent comprising ethanol in the range of 0.10% to 5.0%, and has a pH in the range of 6.0 to 7.4.
25. The process of claim 1 , wherein the process does not comprise centrifugation of the platelet composition, and wherein the TFF-treated composition comprises at least 2000×10 3 platelets/μl in the aqueous medium, and 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.