Control of trace metals during production of anti-CD38 antibodies
The disclosure relates to methods to control trace metals during production of anti-CD38 antibodies, drug substances and drug products generated using the methods, and uses of the generated drug substances and drug products.
1. A method of producing an anti-CD38 antibody expressed from a polynucleotide encoding a heavy chain variable region (VH) of SEQ ID NO: 7 and a light chain variable region (VL) of SEQ ID NO: 8 having an asialo, mono-galacto core-fucosylated biantennary glycan (G1F) oligosaccharide content between 15% and 27%, comprising:
a) preparing a culture medium comprising between 2 parts per billion (ppb) and 8.5 manganese (Mn); and
b) controlling the G1F oligosaccharide content of the anti-CD38 antibody by culturing a host cell comprising the polynucleotide encoding the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8 in the culture medium prepared in step a), thereby producing the anti-CD38 antibody expressed from the polynucleotide encoding the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8 having the G1F oligosaccharide content between 15% and 27%.
2. The method of claim 1 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 25%.
3. The method of claim 2 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 21% and 25%.
4. The method of claim 1 , wherein an asialo, agalacto core-fucosylated biantennary glycan (G0F) oligosaccharide content of the anti-CD38 antibody is between 65% and 74%.
5. The method of claim 4 , wherein the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%.
6. The method of claim 1 , wherein preparing the culture medium comprises:
measuring Mn concentration in one or more batches of raw material components,
selecting those one or more batches of raw material components that in combination contain between 2 ppb and 8.5 ppb Mn, and
using the selected one or more batches of raw material components to prepare the culture medium.
7. The method of claim 6 , wherein the culture medium is prepared to comprise between 4.0 ppb and 8.5 ppb Mn.
8. The method of claim 7 , wherein the culture medium is prepared to comprise between 4.0 ppb and 6.5 ppb Mn.
9. The method of claim 8 , wherein the culture medium is prepared to comprise between 5.0 ppb and 6.5 ppb Mn.
10. The method of claim 1 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 27%, the G0F oligosaccharide content of the anti-CD38 antibody is between 65% and 74%, and the culture medium is prepared to comprise between 4.0 ppb and 8.5 ppb Mn.
11. The method of claim 1 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 25%, the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%, and the culture medium is prepared to comprise between 4.0 ppb and 6.5 ppb Mn.
12. The method of claim 11 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 21% and 25%, the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%, and the culture medium is prepared to comprise between 5.0 ppb and 6.5 ppb Mn.
13. The method of claim 1 , wherein the culture medium is a basal medium or a feed medium.
14. The method of claim 13 , wherein culturing comprises a fed-batch culture or a perfusion culture.
15. The method of claim 14 , wherein the host cell is an eukaryotic cell.
16. The method of claim 15 , wherein the eukaryotic cell is a CHO cell, a PER.C6 cell, a NS0 cell, a Sp2/0 cell or a BHK cell.
17. The method of claim 16 , wherein the CHO cell is a CHO-K1 cell, a CHO-DG44 cell, a CHO-S cell or a CHO-DXB11 cell.
18. The method of claim 17 , wherein the CHO cell is deficient in glutamine synthetase (GS).
19. The method of claim 1 , wherein the method is conducted under good manufacturing practice (GMP) compliant conditions.
20. The method of claim 19 , wherein the anti-CD38 antibody comprises the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8.
21. The method of claim 20 , wherein the anti-CD38 antibody comprises an IgG1 isotype.
22. The method of claim 21 , wherein the anti-CD38 antibody comprises a heavy chain (HC) of SEQ ID NO: 9 and a light chain (LC) of SEQ ID NO: 10.
23. The method of claim 1 , wherein the anti-CD38 antibody is a biosimilar.
24. The method of claim 1 , wherein the culture medium is prepared to comprise between 2 ppb and 6.5 ppb Mn.
25. The method of claim 1 , wherein the anti-CD38 antibody is daratumumab.
26. A method of producing an anti-CD38 antibody expressed from a polynucleotide encoding the VH of SEQ ID NO: 7 and a VL of SEQ ID NO: 8 having a G1F oligosaccharide content between 15% and 27%, comprising:
a) culturing a host cell expressing the polynucleotide encoding the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8 in conditions that the anti-CD38 antibody is produced; and
b) controlling the G1F oligosaccharide content of the anti-CD38 antibody by monitoring the concentration of Mn in the culture medium during biosynthesis of the anti-CD38 antibody and regulating the concentration of Mn in the culture medium during biosynthesis of the anti-CD38 antibody, wherein the concentration of Mn in the culture medium is regulated to comprise between 2 ppb and 8.5 ppb Mn, thereby producing the anti-CD38 antibody having the G1F oligosaccharide content between 15% and 27%.
27. The method of claim 26 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 25%.
28. The method of claim 27 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 21% and 25%.
29. The method of claim 26 , wherein a G0F oligosaccharide content of the anti-CD38 antibody is between 65% and 74%.
30. The method of claim 27 , wherein the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%.
31. The method of claim 26 , wherein the concentration of Mn in the culture medium is regulated to comprise between 4.0 ppb and 8.5 ppb Mn.
32. The method of claim 31 , wherein the concentration of Mn in the culture medium is regulated to comprise between 4.0 ppb and 6.5 ppb Mn.
33. The method of claim 32 , wherein the concentration of Mn in the culture medium is regulated to comprise between 5.0 ppb and 6.5 ppb Mn.
34. The method of claim 26 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 27%, the G0F oligosaccharide content of the anti-CD38 antibody is between 65% and 74%, and the culture medium is regulated to comprise between 4.0 ppb and 8.5 ppb Mn.
35. The method of claim 27 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 25%, the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%, and the culture medium is regulated to comprise between 4.0 ppb and 6.5 ppb Mn.
36. The method of claim 28 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 21% and 25%, the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%, and the culture medium is regulated to comprise between 5.0 ppb and 6.5 ppb Mn.
37. The method of claim 26 , wherein the culture medium is a basal medium or a feed medium.
38. The method of claim 37 , wherein culturing comprises a fed-batch culture or a perfusion culture.
39. The method of claim 38 , wherein the host cell is an eukaryotic cell.
40. The method of claim 39 , wherein the eukaryotic cell is a CHO cell, a PER.C6 cell, a NS0 cell, a Sp2/0 cell or a BHK cell.
41. The method of claim 40 , wherein the CHO cell is a CHO-K1 cell, a CHO-DG44 cell, a CHO-S cell or a CHO-DXB11 cell.
42. The method of claim 41 , wherein the CHO cell is deficient in GS.
43. The method of claim 26 , wherein the method is conducted under GMP-compliant conditions.
44. The method of claim 43 , wherein the anti-CD38 antibody comprises the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8.
45. The method of claim 44 , wherein the anti-CD38 antibody comprises an IgG1 isotype.
46. The method of claim 45 , wherein the anti-CD38 antibody comprises the HC of SEQ ID NO: 9 and the LC of SEQ ID NO: 10.
47. The method of claim 26 , wherein the anti-CD38 antibody is a biosimilar.
48. The method of claim 26 , wherein the culture medium is regulated to comprise between 2 ppb and 6.5 ppb Mn.
49. The method of claim 26 , wherein the anti-CD38 antibody is daratumumab.
50. A method of producing a drug product comprising an anti-CD38 antibody expressed from a polynucleotide encoding the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8 and having a G1F oligosaccharide content between 15% and 27%, comprising:
a) preparing a culture medium comprising between 2 ppb and 8.5 ppb Mn;
b) controlling the G1F oligosaccharide content of the anti-CD38 antibody by culturing a host cell comprising the polynucleotide encoding the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8 in the culture medium prepared in step a), thereby producing the anti-CD38 antibody having the G1F oligosaccharide content between 15% and 27%; and
c) formulating the anti-CD38 antibody as a drug product.
51. The method of claim 50 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 25%.
52. The method of claim 51 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 21% and 25%.
53. The method of claim 50 , wherein a G0F oligosaccharide content of the anti-CD38 antibody is between 65% and 74%.
54. The method of claim 53 , wherein the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%.
55. The method of claim 50 , wherein preparing the culture medium comprises:
measuring Mn concentration in one or more batches of raw material components,
selecting those one or more batches of raw material components that in combination contain between 2 ppb and 8.5 ppb Mn, and
using the selected one or more batches of raw material components to prepare the culture medium.
56. The method of claim 50 , wherein the culture medium is prepared to comprise between 4.0 ppb and 8.5 ppb Mn.
57. The method of claim 56 , wherein the culture medium is prepared to comprise between 4.0 ppb and 6.5 ppb Mn.
58. The method of claim 57 , wherein the culture medium is prepared to comprise between 5.0 ppb and 6.5 ppb Mn.
59. The method of claim 50 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 27%, the G0F oligosaccharide content of the anti-CD38 antibody is between 65% and 74%, and the culture medium is prepared to comprise between 4.0 ppb and 8.5 ppb Mn.
60. The method of claim 59 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 15% and 25%, the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%, and the culture medium is prepared to comprise between 4.0 ppb and 6.5 ppb Mn.
61. The method of claim 60 , wherein the G1F oligosaccharide content of the anti-CD38 antibody is between 21% and 25%, the G0F oligosaccharide content of the anti-CD38 antibody is between 68% and 74%, and the culture medium is prepared to comprise between 5.0 ppb and 6.5 ppb Mn.
62. The method of claim 50 , wherein the culture medium is a basal medium or a feed medium.
63. The method of claim 62 , wherein culturing comprises a fed-batch culture or a perfusion culture.
64. The method of claim 63 , wherein the host cell is an eukaryotic cell.
65. The method of claim 64 , wherein the eukaryotic cell is a CHO cell, a PER.C6 cell, a NS0 cell, a Sp2/0 cell or a BHK cell.
66. The method of claim 65 , wherein the CHO cell is a CHO-K1 cell, a CHO-DG44 cell, a CHO-S cell or a CHO-DXB11 cell.
67. The method of claim 66 , wherein the CHO cell is deficient in glutamine synthetase (GS).
68. The method of claim 50 , wherein the method is conducted under GMP-compliant conditions.
69. The method of claim 68 , wherein the anti-CD38 antibody comprises the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8.
70. The method of claim 69 , wherein the anti-CD38 antibody comprises an IgG1 isotype.
71. The method of claim 70 , wherein the anti-CD38 antibody comprises the HC of SEQ ID NO: 9 and the LC of SEQ ID NO: 10.
72. The method of claim 50 , wherein the anti-CD38 antibody is a biosimilar.
73. The method of claim 50 , wherein formulating the drug product comprises formulating the anti-CD38 antibody at from 20 mg/mL to 180 mg/mL with recombinant human hyaluronidase (rHuPH20) in an amount of from 30,000 U to 45,000 U, histidine at a concentration of from 5 mM to 15 mM, sorbitol at a concentration of from 100 mM to 300 mM, PS-20 at a concentration of from 0.01% w/v to 0.04% w/v and methionine at a concentration of from 1 mg/mL to 2 mg/mL, at a pH of 5.0 to 6.0.
74. The method of claim 73 , wherein formulating the drug product comprises formulating the anti-CD38 antibody at 120 mg/mL in 2,000 U/ml recombinant human hyaluronidase (rHuPH20), 5 mM to 15 mM histidine, 100 mM to 300 mM sorbitol, 0.01% w/v to 0.04% w/v PS-20 and 1 mg/mL to 2 mg/mL methionine, at a pH of 5.6.
75. The method of claim 50 , wherein formulating the drug product comprises formulating the anti-CD38 antibody at 20 mg/ml in 25 mM acetic acid, 60 mM sodium chloride, 140 mM mannitol and 0.04% w/v polysorbate-20 (PS-20); at pH 5.5.
76. The method of claim 50 , wherein the culture medium is prepared to comprise between 2 ppb and 6.5 ppb Mn.
77. The method of claim 50 , wherein the anti-CD38 antibody is daratumumab.