Stabilized liquid anti-RSV antibody formulations
The present invention provides liquid formulations of SYNAGIS® or an antigen-binding fragment thereof that immunospecifically bind to a respiratory syncytial virus (RSV) antigen, which formulation exhibit stability, low to undetectable levels of aggregation, and very little to no loss of the biological activities of SYNAGIS® or an antigen-binding fragment thereof, even during long periods of storage. In particular, the present invention provides liquid formulations of SYNAGIS® or an antigen-binding fragment thereof which immunospecifically binds to a RSV antigen, which formulation are substantially free of surfactant, inorganic salts, and/or other common excipients. Furthermore, the invention provides method of preventing, treating or ameliorating symptoms associated with RSV infection utilizing liquid formulations of the present invention.
1. A method of purifying and formulating a liquid palivizumab formulation comprising at least 15 mg/ml of palivizumab, or an antigen-binding fragment thereof, and histidine in an aqueous carrier, wherein the histidine concentration is about 1 mM to about 100 mM and wherein said formulation is substantially free of surfactant, inorganic salts or other excipients;
wherein the method comprises maintaining the palivizumab, or an antigen-binding fragment thereof, in an aqueous solution at all times during preparation.
2. A method of preparing a liquid formulation of palivizumab, or an antigen-binding fragment thereof, the method comprising:
diafiltering a preparation of palivizumab, or an antigen-binding fragment thereof, into a formulation buffer to obtain a liquid formulation of palivizumab, or an antigen-binding fragment thereof;
wherein the preparation of palivizumab, or an antigen-binding fragment thereof, comprises about 15 mg/ml to about 300 mg/ml palivizumab, or an antigen-binding fragment thereof;
wherein the formulation buffer comprises about 1 mM to about 100 mM histidine;
wherein the method comprises maintaining the palivizumab, or an antigen-binding fragment thereof, in an aqueous phase at all times during preparation;
wherein the liquid formulation of palivizumab, or an antigen-binding fragment thereof, is substantially free of surfactant, inorganic salts and/or other excipients.
3. The method of claim 2 , wherein the method does not involve a drying step.
4. The method of claim 2 , wherein the formulation buffer comprises histidine at a concentration ranging from about 10 mM to about 50 mM histidine, about 20 mM to about 30 mM histidine, about 23 mM to about 27 mM histidine, or about 25 mM histidine.
5. The method of claim 2 , wherein the formulation buffer further comprises glycine at a concentration of less than 150 mM glycine, less than 100 mM glycine, less than 50 mM glycine, less than 3.0 mM glycine, less than 2.0 mM glycine, less than 1.8 mM glycine, or about 1.6 mM glycine.
6. The method of claim 2 , wherein the formulation buffer comprises a pH ranging from about 5.0 to about 7.0, about 5.5 to about 6.5, about 5.8 to about 6.2, or about 6.0.
7. The method of claim 2 , wherein the formulation buffer comprises 25 mM histidine, 1.6 mM glycine, and pH 6.
8. The method of claim 2 , comprising concentrating the preparation of palivizumab, or an antigen-binding fragment thereof, to a concentration of about 15 mg/ml to about 300 mg/ml prior to diafiltration.
9. The method of claim 8 , wherein the preparation of palivizumab, or an antigen-binding fragment thereof, is concentrated to a concentration of about 15 mg/ml, about 20 mg/ml, about 30 mg/ml, about 40 mg/ml, about 50 mg/ml, about 60 mg/ml, about 70 mg/ml, about 80 mg/ml, about 90 mg/ml, about 100 mg/ml, about 150 mg/ml, about 200 mg/ml, about 250 mg/ml, or about 300 mg/ml prior to diafiltration.
10. The method of claim 2 , wherein the liquid formulation of palivizumab, or an antigen-binding fragment thereof, is diluted to a concentration of 103±3 mg/ml palivizumab, or an antigen-binding fragment thereof.
11. The method of claim 2 , further comprising sterilization of the liquid formulation.
12. The method of claim 11 , wherein sterilization comprises sterile filtration or radiation.
13. The method of claim 11 , wherein sterilization comprises filter sterilization comprising a 0.2 or 0.22 micron filter.
14. The method of claim 2 , comprising purifying the preparation of palivizumab, or an antigen-binding fragment thereof, prior to diafiltration.
15. The method of claim 14 , wherein purifying the preparation of palivizumab, or an antigen-binding fragment thereof, comprises:
HS50 chromatography;
benzonase treatment;
Protein A chromatography;
nanofiltration;
low pH treatment; and/or
Q chromatography column cycle.
16. The method of claim 2 , wherein the preparation of palivizumab, or an antigen-binding fragment thereof, is purified from conditioned cell culture medium.
17. The method of claim 16 , wherein purifying palivizumab, or an antigen-binding fragment thereof, from conditioned cell culture medium comprises ion exchange chromatography, affinity chromatography, Protein A chromatography, antigen affinity chromatography, and/or sizing column chromatography.
18. The method of claim 10 , wherein the formulation buffer comprises 25 mM histidine, 1.6 mM glycine, and pH 6.
19. The method of 1 , wherein the method does not involve a drying step.
20. The method of claim 1 , wherein the formulation buffer comprises histidine at a concentration ranging from about 10 mM to about 50 mM histidine, about 20 mM to about 30 mM histidine, about 23 mM to about 27 mM histidine, or about 25 mM histidine.
21. The method of claim 1 , wherein the formulation buffer further comprises glycine at a concentration of less than 150 mM glycine, less than 100 mM glycine, less than 50 mM glycine, less than 3.0 mM glycine, less than 2.0 mM glycine, less than 1.8 mM glycine, or about 1.6 mM glycine.
22. The method of claim 1 , wherein the formulation buffer comprises a pH ranging from about 5.0 to about 7.0, about 5.5 to about 6.5, about 5.8 to about 6.2, or about 6.0.
23. The method of claim 1 , wherein the formulation buffer comprises 25 mM histidine, 1.6 mM glycine, and pH 6.
24. The method of claim 1 , wherein the liquid palivizumab formulation or an antigen binding fragment thereof comprises about 15 mg/ml, about 20 mg/ml, about 30 mg/ml, about 40 mg/ml, about 50 mg/ml, about 60 mg/ml, about 70 mg/ml, about 80 mg/ml, about 90 mg/ml, about 100 mg/ml, about 150 mg/ml, about 200 mg/ml, about 250 mg/ml, or about 300 mg/ml of palivizumab.
25. The method of claim 1 , wherein the liquid formulation of palivizumab or an antigen-binding fragment thereof, comprises 103±3 mg/ml palivizumab, or an antigen-binding fragment thereof.
26. The method of claim 25 , wherein the formulation buffer comprises 25 mM histidine, 1.6 mM glycine, and pH 6.
27. The method of claim 1 , further comprising sterilization of the liquid formulation.
28. The method of claim 27 , wherein sterilization comprises sterile filtration or radiation.
29. The method of claim 27 , wherein sterilization comprises filter sterilization comprising a 0.2 or 0.22 micron filter.
30. The method of claim 1 , wherein purifying the preparation of palivizumab, or an antigen-binding fragment thereof, comprises:
HS50 chromatography;
benzonase treatment;
Protein A chromatography;
nanofiltration;
low pH treatment; and/or
Q chromatography column cycle.
31. The method of claim 1 , wherein the preparation of palivizumab, or an antigen-binding fragment thereof, is purified from conditioned cell culture medium.
32. The method of claim 31 , wherein purifying palivizumab, or an antigen-binding fragment thereof, from conditioned cell culture medium comprises ion exchange chromatography, affinity chromatography, Protein A chromatography, antigen affinity chromatography, and/or sizing column chromatography.