Method to produce a highly concentrated immunoglobulin preparation for subcutaneous use
View Patent ↗The present invention relates to a new and improved method for preparing a highly concentrated immunoglobulin composition from pooled plasma for subcutaneous injection. A composition comprising 20% or more immunoglobulin suitable for subcutaneous use is also described.
1. A method for preparing a concentrated immunoglobulin G (IgG) composition, comprising the steps:
(A) concentrating a first solution comprising IgG to a protein concentration of from 2% to 10% (w/v) by ultrafiltration using a first ultra-/diafiltration system comprising a first ultrafiltration membrane, thereby forming a first IgG concentrate;
(B) diafiltering the first IgG concentrate against a diafiltration buffer using the first ultra-diafiltration system comprising the first ultrafiltration membrane, thereby forming a first IgG diafiltrate;
(C) concentrating the first IgG diafiltrate to a protein concentration of greater than 20% (w/v) by ultrafiltration using the first ultra-/diafiltration system comprising the first ultrafiltration membrane, thereby forming a second IgG concentrate;
(D) collecting the second IgG concentrate from the first ultra-/diafiltration system;
(E) washing the first ultrafiltration membrane by re-circulating a post-wash buffer through the first ultra-/diafiltration system wherein the first ultra-/diafiltration system is washed with a volume of post-wash buffer equal to at least two times the dead volume of the first ultra-/diafiltration system, thereby forming a first IgG post-wash solution;
(F) transfering the first IgG post-wash solution from the first ultra-/diafiltration system into a second ultra-/diafiltration system comprising a second ultrafiltration membrane, wherein the surface area of the second ultrafiltration membrane is lower than the surface area of the first ultrafiltration membrane;
(G) concentrating the first IgG post-wash solution to a protein concentration of greater than 20% (w/v) by ultrafiltration using the second ultra-/diafiltration system comprising a second ultra-/diafiltration membrane, thereby forming a third IgG concentrate; and
(H) combining the third IgG concentrate from the second ultra-/diafiltration system with the second IgG concentrate, thereby forming a concentrated IgG composition.
2. The method of claim 1 , wherein the first solution comprising IgG is concentrated in (A) to a protein concentration of 5±1% (w/v).
3. The method of claim 1 , wherein the first ultrafiltation membrane is an open-screen membrane.
4. The method of claim 1 , wherein the first or second ultrafiltration membrane have a nominal molecular weight cut off (NMWCO) of 100 kDa or less.
5. The method of claim 1 , wherein the first and second ultrafiltration membrane has a nominal molecular weight cut off (NMWCO) of 100 kDa or less.
6. The method of claim 1 , wherein the first or second ultrafiltration membrane has a nominal molecular weight cut off (NMWCO) of 90 kDa or less.
7. The method of claim 1 , wherein the first and second ultrafiltration membrane have a nominal molecular weight cut off (NMWCO) of 90 kDa or less.
8. The method of claim 1 , wherein the first and second ultrafiltration membrane have a same nominal molecular weight cut off (NMWCO).
9. The method of claim 1 , wherein the diafiltration buffer comprises from 0.2 M to 0.3 M glycine and a pH of 4.2±0.1.
10. The method of claim 1 , wherein the second IgG concentrate has a protein concentration of at least 22% (w/v).
11. The method of claim 1 , wherein the surface area of the second ultrafiltration membrane is no more than a tenth of the surface area of the first ultrafiltration membrane.
12. The method of claim 1 , wherein the protein concentration of the concentrated IgG composition formed in (H) is greater than 20% (w/v), and wherein the method further comprises adjusting the concentration of the concentrated IgG composition formed in (H) to about 20% (w/v).
13. The method of claim 1 , wherein the pH of the concentrated IgG composition formed in (H) is from 4.0 to 6.0.
14. The method of claim 1 , further comprising the steps of:
(I) washing the second ultrafiltration membrane by re-circulating a post-wash buffer through the second ultra-/diafiltration system, thereby forming a second IgG post-wash solution; and
(J) adjusting the protein concentration of the concentrated IgG composition formed in (H) by admixing at least a portion of the second IgG post-wash solution into the concentrated IgG composition formed in (H).
15. The method of claim 1 , wherein the IgG is human IgG.
16. The method of claim 1 , comprising the steps:
(A) concentrating a first solution comprising IgG to a protein concentration of 5±1% (w/v) by ultrafiltration using a first ultra-/diafiltration system comprising a first ultrafiltration membrane, thereby forming a first IgG concentrate, the first ultrafiltration membrane having a NMWCO of 60 kDa or less;
(B) diafiltering the first IgG concentrate against a diafiltration buffer using the first ultra-/diafiltration system comprising the first ultrafiltration membrane, thereby forming a first IgG diafiltrate, the diafiltration buffer comprising glycine and a pH of 4.2±0.1;
(C) concentrating the first IgG diafiltrate to a protein concentration of greater than 20% (w/v) by ultrafiltration using the first ultra-/diafiltration system comprising the first ultrafiltration membrane, thereby forming a second IgG concentrate;
(D) collecting the second IgG concentrate from the first ultra-/diafiltration system;
(E) washing the first ultrafiltration membrane by re-circulating a post-wash buffer through the first ultra-/diafiltration system, thereby forming a first IgG post-wash solution, the post-wash buffer having a volume equal to at least two times the dead volume of the first ultra-/diafiltration system;
(F) collecting the first IgG post-wash solution from the first ultra-/diafiltration system into a second ultra-/diafiltration system comprising a second ultrafiltration membrane, the second ultrafiltration membrane having a NMWCO of 60 kDa or less;
(G) concentrating the first IgG post-wash solution to a protein concentration of greater than 20% (w/v) by ultrafiltration using the second ultra-/diafiltration system comprising a second ultra-/diafiltration membrane, wherein the surface area of the second ultrafiltration membrane is no more than a tenth of the surface area of the first ultrafiltration membrane, thereby forming a third IgG concentrate;
(H) combining the third IgG concentrate from the second ultra-/diafiltration system with the second IgG concentrate, thereby forming a concentrated IgG composition;
(I) washing the second ultrafiltration membrane by re-circulating a post-wash buffer through the second ultra-/diafiltration system, thereby forming a second IgG post-wash solution; and
(J) adjusting the protein concentration of the concentrated IgG composition formed in (H) by admixing at least a portion of the second IgG post-wash solution into the concentrated IgG composition formed in (H).