Method to produce an immunoglobulin preparation with improved yield
The present invention provides improved methods for the manufacturing of IVIG products. These methods offer various advantages such as reduced loss of IgG during purification and improved quality of final products. In other aspects, the present invention provides aqueous and pharmaceutical compositions suitable for intravenous, subcutaneous, and/or intramuscular administration. In yet other embodiments, the present invention provides methods of treating a disease or condition comprising administration of an IgG composition provided herein.
1. A method for preparing an enriched IgG composition from plasma, the method comprising the steps of:
(a) precipitating a cryo-poor plasma fraction, in a first precipitation step, with from about 6% to about 10% alcohol at a pH of from about 7.0 to about 7.5 to obtain a first precipitate and a first supernatant;
(b) precipitating IgG from the first supernatant, in a second precipitation step, with from about 23% to about 27% alcohol at a pH of from about 6.7 to about 7.1 at a temperature of from −7° C. to −9° C. to form a second precipitate;
(c) suspending the second precipitate to form a suspension;
(d) precipitating IgG from the suspension formed in step (c), in a third precipitation step, with from 22% to 28% alcohol at a pH of from 6.7 to 7.3 to form a third precipitate;
(e) re-suspending the third precipitate to form a suspension; and
(f) separating the soluble fraction from the suspension formed in step (e), thereby forming an enriched IgG composition.
2. The method of claim 1 , wherein the temperature of precipitation step (b) is −7° C.
3. The method of claim 1 , wherein the alcohol concentration of precipitation step (b) is about 25%.
4. The method of claim 2 , wherein the alcohol concentration of precipitation step (b) is about 25%.
5. The method of claim 1 , wherein at least one of the first precipitation step, second precipitation step, or third precipitation step comprises spray addition of the alcohol, the spray addition in the form of fine droplets or mist of the alcohol.
6. The method of claim 1 , wherein the first precipitation step, second precipitation step, and third precipitation step comprise spray addition of the alcohol, the spray addition in the form of fine droplets or mist of the alcohol.
7. The method of claim 1 , wherein the pH of at least one of the first precipitation step, second precipitation step, or third precipitation step is achieved by addition of a pH modifying solution after addition of the alcohol.
8. The method of claim 7 , wherein the addition of a pH modifying solution comprises spray addition of the pH modifying solution.
9. The method of claim 1 , wherein the pH of all of the first precipitation step, second precipitation step, and third precipitation step is achieved by addition of a pH modifying solution after addition of the alcohol.
10. The method of claim 9 , wherein the addition of a pH modifying solution comprises spray addition of the pH modifying solution.
11. The method of claim 1 , wherein the pH of the precipitation step is modified before and after the addition of alcohol, during and after the addition of alcohol, or before, during, and after the addition of alcohol.
12. The method of claim 1 , wherein the pH of at least one precipitation step is maintained for the entire precipitation step by continuous adjustment of the pH.
13. The method of claim 1 , wherein the precipitate formed in step (b) is suspended with from about 12 L to about 18 L of buffer per kg precipitate.
14. The method of claim 1 , wherein the precipitate formed in step (b) is suspended with about 15 L of buffer per kg precipitate.
15. The method of claim 1 , wherein the suspension formed in step (c) is treated with finely divided silicon dioxide.
16. The method of claim 15 , wherein the suspension formed in step (c) is treated with from about 0.02 kg to about 0.06 kg of finely divided silicon dioxide per kg of the second precipitate suspended.
17. The method of claim 15 , wherein treatment with finely divided silicon dioxide comprises mixing the finely divided silicon dioxide with the suspension formed in step (c) for at least 30 minutes.
18. The method of claim 17 , wherein the suspension formed in step (c) is treated with from about 0.02 kg to about 0.06 kg of finely divided silicon dioxide per kg of the second precipitate suspended.
19. The method of claim 1 , wherein the method further comprises an ion exchange chromatography purification step.
20. The method of claim 1 , wherein the method further comprises a nanofiltration step.
21. The method of claim 20 , wherein the nanofiltration step comprises the use of a nanofilter having a mean pore size of about 35 nm.
22. The method of claim 1 , wherein the enriched IgG composition obtained in step (f) contains at least 85% of the IgG content found in the cryo-poor plasma fraction used in step (a).
23. The method of claim 1 , wherein the enriched IgG composition obtained in step (f) contains at least 90% of the IgG content found in the cryo-poor plasma fraction used in step (a).
24. The method of claim 1 , wherein the purity of IgG in the final composition is at least 95%.
25. The method of claim 1 , wherein the purity of IgG in the final composition is at least 98%.
26. The method of claim 1 , wherein the purity of IgG in the final IgG composition is at least 99%.
27. The method of claim 1 , wherein the plasma is human plasma.