Crystals of whole antibodies and fragments thereof and methods for making and using them
View Patent ↗This invention relates to crystals of whole antibodies and fragments thereof, and formulations and compositions comprising such crystals. More particularly, methods are provided for the crystallization of high concentrations of whole antibodies, and fragments thereof, in large batches, and for the preparation of stabilized whole antibody crystals for use alone, or in dry or slurry formulations or compositions. This invention also relates to methods for stabilization, storage and delivery of biologically active whole antibody crystals.
1. A crystal of infliximab, wherein the crystal comprises infliximab, ethoxyethanol, lithium sulfate, and Tris buffer.
2. The crystal of claim 1 , wherein the pH of the Tris buffer is 8.6.
3. A method of crystallizing infliximab, the method comprising:
combining infliximab, ethoxyethanol, lithium sulfate, and Tris buffer, thereby forming a crystallization solution; and
incubating the crystallization solution, thereby crystallizing infliximab.
4. The method of claim 3 , wherein the method is performed at room temperature.
5. The method of claim 3 , wherein the method is performed at pH 8.6.
6. The method of claim 3 , wherein the concentration of infliximab in the crystallization solution is
2.5
0.15
mg
/
ml
.
7. The method of claim 3 , wherein the percentage of ethoxyethanol in the crystallization solution is
3500
150
%
.
8. The method of claim 3 , wherein the concentration of lithium sulfate in the crystallization solution is
20
150
M
.
9. The method of claim 3 , wherein the concentration of Tris buffer in the crystallization solution is
10
150
M
.
10. The method of claim 3 , wherein the concentration of infliximab in the crystallization solution is
2.5
0.15
mg
/
ml
,
the percentage of ethoxyethanol in the crystallization solution is
3500
150
%
,
the concentration of lithium sulfate in the crystallization solution is
20
150
M
,
and the concentration of Tris buffer in the crystallization solution is
10
150
M
.
11. A crystal of infliximab, wherein the crystal comprises infliximab, PEG-400, lithium sulfate, and Tris buffer.
12. The crystal of claim 11 , wherein the pH of the Tris buffer is 8.5.
13. A method of crystallizing infliximab, the method comprising:
combining infliximab, PEG-400, lithium sulfate, and Tris buffer, thereby forming a crystallization solution; and
incubating the crystallization solution, thereby crystallizing infliximab.
14. The method of claim 13 , wherein the method is performed at room temperature.
15. The method of claim 13 , wherein the method is performed at pH 8.5.
16. The method of claim 13 , wherein the concentration of infliximab in the crystallization solution is
2.5
0.15
mg
/
ml
.
17. The method of claim 13 , wherein the percentage of PEG-400 in the crystallization solution is
4000
150
%
.
18. The method of claim 13 , wherein the concentration of lithium sulfate in the crystallization solution is
20
150
M
.
19. The method of claim 13 , wherein the concentration of Tris buffer in the crystallization solution is
10
150
M
.
20. The method of claim 13 , wherein the concentration of infliximab in the crystallization solution is
2.5
0.15
mg
/
ml
,
the percentage of PEG-400 in the crystallization solution is
4000
150
%
,
the concentration of lithium sulfate in the crystallization solution is
20
150
M
,
and the concentration of Tris buffer in the crystallization solution is
10
150
M
.
21. A crystal of infliximab, wherein the crystal comprises infliximab, polyethylene glycol monomethyl ether 550 (PEG MME 550), calcium chloride, and Tris HCl buffer.
22. The crystal of claim 21 , wherein the pH of the Tris HCl buffer is 7.0.
23. A method of crystallizing infliximab, the method comprising:
combining infliximab, PEG MME 550, calcium chloride, and Tris HCl buffer, thereby forming a crystallization solution; and
incubating the crystallization solution, thereby crystallizing infliximab.
24. The method of claim 23 , wherein the method is performed at room temperature.
25. The method of claim 23 , wherein the pH of the Tris HCl buffer is 7 . 0 .
26. The method of claim 23 , wherein the concentration of infliximab in the crystallization solution is
1.25
0.033
mg
/
ml
.
27. The method of claim 23 , wherein the percentage of PEG MME 550 in the crystallization solution is
500
33
%
.
28. The method of claim 23 , wherein the concentration of calcium chloride in the crystallization solution is
3
33
M
.
29. The method of claim 23 , wherein the concentration of Tris HCl buffer in the crystallization solution is
2.5
33
M
.
30. The method of claim 23 , wherein the concentration of infliximab in the crystallization solution is
1.25
0.033
mg
/
ml
,
the percentage of PEG MME 550 in the crystallization solution is
500
33
%
,
the concentration of calcium chloride in the crystallization solution is
3
33
M
,
and the concentration of Tris HCl buffer in the crystallization solution is
2.5
33
M
.
31. A crystal of infliximab, wherein the crystal comprises infliximab, PEG 300, Tris buffer, PEG 8000, and glycerol.
32. The crystal of claim 31 , wherein the pH of the Tris buffer is 8.5.
33. A method of crystallizing infliximab, the method comprising:
combining infliximab, PEG 300, Tris buffer, PEG 8000, and glycerol, thereby forming a crystallization solution; and
incubating the crystallization solution, thereby crystallizing infliximab.
34. The method of claim 33 , wherein the method is performed at room temperature.
35. The method of claim 33 , wherein the pH is 8.5.
36. The method of claim 33 , wherein the concentration of infliximab in the crystallization solution is
0.5
0.075
mg
/
ml
.
37. The method of claim 33 , wherein the percentage of PEG 300 in the crystallization solution is
1000
75
%
.
38. The method of claim 33 , wherein the concentration of Tris buffer in the crystallization solution is
5
75
M
.
39. The method of claim 33 , wherein the percentage of PEG 8000 in the crystallization solution is
250
75
%
.
40. The method of claim 33 , wherein the percentage of glycerol in the crystallization solution is
500
75
%
.
41. The method of claim 33 , wherein the concentration of infliximab in the crystallization solution is
0.5
0.075
mg
/
ml
,
the percentage of PEG 300 in the crystallization solution is
1000
75
%
,
the concentration of Tris buffer in the crystallization solution is
5
75
M
,
the percentage of PEG 8000 in the crystallization solution is
250
75
%
,
and the percentage of glycerol in the crystallization solution is
500
75
%
.