Antibody formulations and methods of making same

• US 8,420,081 B2
• Filed: 11/28/2008
• Issued: 04/16/2013
• Est. Priority Date: 11/30/2007
• Status: Active Grant

• Current Assignee: AbbVie Biotechnology Ltd. (Puerto Rico) (Abbvie Incorporated)
• Original Assignee: Abbvie Incorporated
• Inventors: Fraunhofer, Wolfgang, Krause, Hans-juergen, Tschoepe, Markus, Kaleta, Katharina, Bartl, Annika
• Primary Examiner(s): Landsman, Robert
• Assistant Examiner(s): Hissong, Bruce D
• Application Number:
  US12/325,049
• Publication Number:
  US 20090291062A1
• Time in Patent Office: 1,600 Days
• Field of Search: None
• US Class Current: 424/130.1
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 38/46 Hydrolases (3)
• CPC Class Codes: A61K 38/4893 Botulinum neurotoxin (3.4.24.69)
• CPC Class Codes: A61K 38/50 acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39591 Stabilisation, fragmentation
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/08 Solutions composition of solutions A61K47/00
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders

Abstract

The invention provides an aqueous formulation comprising water and a protein, and methods of making the same. The aqueous formulation of the invention may be a high protein formulation and/or may have low levels of conductivity resulting from the low levels of ionic excipients. Also included in the invention are formulations comprising water and proteins having low osmolality.

Claims

1. An aqueous formulation comprising an antibody, or antigen-binding fragment thereof, at a concentration of at least about 20 mg/mL and water, wherein the formulation has a conductivity of less than about 2.5 mS/cm and the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

2. The formulation of claim 1, wherein the antibody, or antigen-binding fragment thereof, has a M_w greater than about 57 kDa.

3. The formulation of claim 1, wherein the antibody, or antigen-binding fragment thereof, has a M_w greater than about 100 kDa.

4. The formulation of claim 1, wherein the antibody, or antigen-binding fragment thereof, has a M_w greater than about 150 kDa.

5. The formulation of claim 1, wherein the antibody, or antigen-binding fragment thereof, has a M_w greater than about 250 kDa.

6. The formulation of claim 1, wherein the formulation has a conductivity of less than about 2 mS/cm.

7. The formulation of claim 1, wherein the formulation has a conductivity of less than about 1 mS/cm.

8. The formulation of claim 1, wherein the formulation has a conductivity of less than about 0.5 mS/cm.

9. The formulation of claim 1, wherein the concentration of the antibody, or antigen-binding fragment thereof, is at least about 100 mg/mL.

10. The formulation of claim 1, wherein the concentration of the antibody, or antigen-binding fragment thereof, is at least about 150 mg/mL.

11. The formulation of claim 1, wherein the concentration of the antibody, or antigen-binding fragment thereof, is at least about 200 mg/mL.

12. The formulation of claim 1, wherein the concentration of the antibody, or antigen-binding fragment thereof, is greater than 200 mg/mL.

21. The formulation of any one of claims 1, 13, and 17, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of a chimeric antibody, a human antibody, a humanized antibody, and a domain antibody (dAb).

22. The formulation of any one of claims 1, 13, and 17, wherein the antibody, or antigen-binding fragment thereof, is an anti-tumor necrosis factor alpha (TNFα

) or an anti-interleukin-12 (IL-12) antibody, or antigen-binding fragment thereof.

23. The formulation of claim 22, wherein the anti-IL-12 antibody, or antigen-binding fragment thereof, is ustekinumab.

24. The formulation of claim 22, wherein the anti-TNFα

antibody, or antigen-binding fragment thereof, is selected from the group consisting of infliximab, golimumab, and certolizumab pegol.

25. The formulation of claim 22, wherein the anti-TNFα

antibody, or antigen-binding fragment thereof, is adalimumab.

34. The formulation of any one of claims 1, 13, 17, and 28, further comprising a non-ionizable excipient.

35. The formulation of claim 34, wherein the non-ionizable excipient is selected from the group consisting of a polyol, a non-ionic surfactant, sucrose, trehalose, raffinose, and maltose.

36. The formulation of claim 35, wherein the polyol is mannitol or sorbitol.

37. The formulation of claim 35, wherein the non-ionic surfactant is polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80.

38. The formulation of any one of claims 1, 13, 17, and 28, wherein the formulation is stable in a liquid form for at least about 3 months or at least about 12 months.

39. The formulation of any one of claims 1, 13, 17, and 28, wherein the formulation does not comprise an agent selected from the group consisting of a tonicity modifier, a stabilizing agent, a surfactant, an anti-oxidant, a cryoprotectant, a bulking agent, a lyroprotectant, a basic component, and an acidic component.

40. The formulation of any one of claims 1, 13, 17, and 28, wherein the formulation is suitable for in vitro or in vivo use.

41. The formulation of claim 40, wherein the formulation is suitable for administration to a subject via a mode of administration selected from the group consisting of subcutaneous, intravenous, inhalation, intradermal, transdermal, intraperitoneal, and intramuscular.

42. A device comprising the formulation of any one claims 1, 13, 17, and 28.

43. An article of manufacture comprising the formulation of any one of claims 1, 13, 17, and 28.

13. An aqueous formulation comprising an antibody, or an antigen-binding fragment thereof, at a concentration of at least about 50 mg/mL, and water, wherein the formulation has an osmolality of no more than about 30 mOsmol/kg and wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

14. The formulation of claim 13, wherein the osmolality of the formulation is no more than about 15 mOsmol/kg.

15. The formulation of claim 13, wherein the concentration of the antibody, or an antigen-binding fragment thereof, is at least about 150 mg/mL.

16. The formulation of claim 13, wherein the concentration of the antibody, or an antigen-binding fragment thereof, is greater than about 200 mg/mL.

26. The formulation of claim 13 or 17, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of, alemtuzumab, arcitumomab, cetuximab, trastuzumab, imciromab pentetate, capromab pendetide, infliximab, abciximab, rituximab, basiliximab, palivizumab, nofetumomab, omalizumab, daclizumab, ibritumomab tiuxetan, muromonab-CD3, edrecolomab gemtuzumab ozogamicin, golimumab, certolizumab pegol, eculizumab, ustekinumab, panitumumab, tositumomab, I¹³¹ tositumomab, and bevacizumab.

27. The formulation of claim 13 or 17, wherein the antibody, or antigen-binding fragment thereof, is adalimumab.

17. An aqueous formulation comprising water and at least about 20 mg/mL of an antibody, or an antigen-binding fragment thereof, wherein the antibody, or antigen-binding fragment thereof, has a hydrodynamic diameter (D_h) which is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in a buffered solution at the same concentration, and wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

18. The formulation of claim 17, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in phosphate buffered saline (PBS) at the same concentration.

19. The formulation of claim 18, wherein the D_h of the protein is at least about 60% less than the D_h of the antibody, or antigen-binding fragment thereof, in PBS at the same concentration.

20. The formulation of claim 18, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 70% less than the D_h of the antibody, or antigen-binding fragment thereof, in PBS at the same concentration.

28. An aqueous formulation comprising an antibody, or an antigen-binding fragment, at a concentration of at least about 20 mg/mL, and water, wherein the antibody, or antigen-binding fragment, has a hydrodynamic diameter (D_h) of less than about 4 nm, and wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

29. The formulation of claim 28, wherein the antibody has a D_h of less than about 3 nm.

30. The formulation of claim 28, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of a chimeric antibody, a human antibody, a humanized antibody, and a domain antibody (dAb).

31. The formulation of claim 28, wherein the antibody, or antigen-binding fragment thereof, is an anti-TNF-alpha or an anti-IL-12 antibody, or antigen-binding fragment thereof.

32. The formulation of claim 28, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of, alemtuzumab, arcitumomab, cetuximab, trastuzumab, imciromab pentetate, capromab pendetide, infliximab, abciximab, rituximab, basiliximab, palivizumab, nofetumomab, omalizumab, daclizumab, ibritumomab tiuxetan, muromonab-CD3, edrecolomab gemtuzumab ozogamicin, golimumab, certolizumab pegol, eculizumab, ustekinumab, panitumumab, tositumomab, I¹³¹ tositumomab, and bevacizumab.

33. The formulation of claim 28, wherein the antibody, or antigen-binding fragment thereof, is adalimumab.

44. An aqueous formulation comprising an antibody, or an antigen-binding fragment thereof, at a concentration of at least about 20 mg/ml, the antibody, or antigen-binding fragment thereof, prepared by

a) providing the antibody, or antigen-binding fragment thereof, in a first solution;

and

b) subjecting the first solution to diafiltration using water as a diafiltration medium until at least a five-fold volume exchange with the water has been achieved;

wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

45. An aqueous formulation comprising water and at least about 50 mg/ml of an antibody, or an antigen-binding fragment thereof, wherein the antibody, or antigen-binding fragment thereof, has a hydrodynamic diameter (D_h) which is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in a buffered solution at the same concentration, and wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

46. The formulation of claim 45, wherein the concentration of the antibody, or antigen-binding fragment thereof, is at least about 150 mg/mL.

47. The formulation of claim 45, wherein the concentration of the antibody, or antigen-binding fragment thereof, is greater than 200 mg/mL.

48. The formulation of claim 45, further comprising a non-ionizable excipient.

49. The formulation of claim 48, wherein the non-ionizable excipient is selected from the group consisting of a polyol, a non-ionic surfactant, sucrose, trehalose, raffinose, and maltose.

50. The formulation of claim 49, wherein the non-ionic surfactant is polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80.

51. The formulation of claim 45, wherein the formulation is stable in a liquid form for at least about 3 months or at least about 12 months.

52. The formulation of claim 45, wherein the formulation does not comprise an agent selected from the group consisting of a tonicity modifier, a stabilizing agent, a surfactant, an anti-oxidant, a cryoprotectant, a bulking agent, a lyroprotectant, a basic component, and an acidic component.

53. The formulation of claim 45, wherein the formulation is suitable for in vitro or in vivo use.

54. The formulation of claim 53, wherein the formulation is suitable for administration to a subject via a mode of administration selected from the group consisting of subcutaneous, intravenous, inhalation, intradermal, transdermal, intraperitoneal, and intramuscular.

55. A device comprising the formulation of claim 45.

56. An article of manufacture comprising the formulation of claim 45.

57. An aqueous formulation comprising water and an antibody, or antigen-binding fragment thereof, at a concentration of at least about 50 mg/mL,

wherein the formulation has a conductivity of less than about 2.5 mS/cm, and

wherein the antibody, or antigen-binding fragment thereof, has a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

7, and a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

58. The formulation of claim 57, wherein the concentration of the antibody, or antigen-binding fragment thereof, is at least about 75 mg/mL.

59. The formulation of claim 57, wherein the formulation has a conductivity of less than about 2 mS/cm.

60. The formulation of claim 57, further comprising a non-ionizable excipient.

61. The formulation of claim 60, wherein the non-ionizable excipient is a polyol.

62. The formulation of claim 61, wherein the polyol is mannitol.

63. The formulation of claim 60, wherein the non-ionizable excipient is a non-ionic surfactant.

64. The formulation of claim 63, wherein the non-ionic surfactant is polysorbate 80.

65. The formulation of claim 60, wherein the non-ionizable excipient is sucrose.

66. The formulation of claim 57, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

67. The formulation of claim 57, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

98. A device comprising the formulation of any one of claims 57, 68, or 84.

99. An article of manufacture comprising the formulation of any one of claims 57, 68, or 84.

68. An aqueous formulation comprising water and an antibody, or an antigen-binding fragment thereof, at a concentration of at least about 50 mg/mL,

wherein the antibody, or an antigen-binding fragment thereof, has a hydrodynamic diameter (D_h) which is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in a buffered solution at the same concentration, and

wherein the antibody, or antigen-binding fragment thereof, has a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

7, and a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

69. The formulation of claim 68, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in phosphate buffered saline (PBS) at the same concentration.

70. The formulation of claim 69, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 60% less than the D_h of the antibody, or antigen-binding fragment thereof, in PBS at the same concentration.

71. The formulation of claim 70, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 70% less than the D_h of the antibody, or antigen-binding fragment thereof, in PBS at the same concentration.

72. The formulation of claim 68, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

73. The formulation of claim 68, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

74. The formulation of claim 68, further comprising a non-ionizable excipient.

75. The formulation of claim 74, wherein the non-ionizable excipient is a polyol.

76. The formulation of claim 75, wherein the polyol is mannitol.

77. The formulation of claim 74, wherein the non-ionizable excipient is a non-ionic surfactant.

78. The formulation of claim 77, wherein the non-ionic surfactant is polysorbate 80.

79. The formulation of claim 74, wherein the non-ionizable excipient is sucrose.

80. The formulation of claim 68, further comprising mannitol and polysorbate 80.

81. The formulation of claim 68, further comprising sucrose and polysorbate 80.

82. The formulation of claim 80 or 81, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

83. The formulation of claim 80 or 81, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

84. An aqueous formulation comprising water and an antibody, or an antigen-binding fragment, at a concentration of at least about 50 mg/mL,

wherein the antibody, or antigen-binding fragment, has a hydrodynamic diameter (D_h) of less than about 4 nm, and

wherein the antibody, or antigen-binding fragment thereof, has a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

7, and a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

85. The formulation of claim 84, wherein the antibody, or antigen-binding portion thereof, has a D_h of less than about 3 nm.

86. The formulation of claim 84, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

87. The formulation of claim 84, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

88. The formulation of claim 84, further comprising a non-ionizable excipient.

89. The formulation of claim 88, wherein the non-ionizable excipient is a polyol.

90. The formulation of claim 89, wherein the polyol is mannitol.

91. The formulation of claim 88, wherein the non-ionizable excipient is a non-ionic surfactant.

92. The formulation of claim 91, wherein the non-ionic surfactant is polysorbate 80.

93. The formulation of claim 88, wherein the non-ionizable excipient is sucrose.

94. The formulation of claim 84, further comprising mannitol and polysorbate 80.

95. The formulation of claim 84, further comprising sucrose and polysorbate 80.

96. The formulation of claim 94 or 95 wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

97. The formulation of claim 94 or 95, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

100. An aqueous formulation comprising

mannitol,

polysorbate 80,

an antibody, or antigen-binding fragment thereof, at a concentration of at least about 20 mg/mL, and

water,

wherein the formulation has a conductivity of less than about 2.5 mS/cm, and

wherein the antibody, or antigen-binding fragment thereof, has a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

7, and a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

101. The formulation of claim 100, wherein the formulation has a conductivity of less than about 1.0 mS/cm.

102. The formulation of claim 100, wherein the formulation has a conductivity of less than about 0.5 mS/cm.

103. The formulation of claim 100, wherein the antibody concentration is at least about 50 mg/mL.

104. The formulation of claim 100, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

105. The formulation of claim 100, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

118. A device comprising the formulation of any one of claims 100, 106, and 113.

119. An article of manufacture comprising the formulation of any one of claims 100, 106, and 113.

106. An aqueous formulation comprising

mannitol,

polysorbate 80,

an antibody, or antigen-binding fragment thereof, at a concentration of at least about 20 mg/mL, and

water,

wherein the antibody, or an antigen-binding fragment thereof, has a hydrodynamic diameter (D_h) which is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in a buffered solution at the same concentration, and

wherein the antibody, or antigen-binding fragment thereof, has a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

7, and a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

107. The formulation of claim 106, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 50% less than the D_h of the antibody, or antigen-binding fragment thereof, in phosphate buffered saline (PBS) at the same concentration.

108. The formulation of claim 107, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 60% less than the D_h of the antibody, or antigen-binding fragment thereof, in PBS at the same concentration.

109. The formulation of claim 107, wherein the D_h of the antibody, or antigen-binding fragment thereof, is at least about 70% less than the D_h of the antibody, or antigen-binding fragment thereof, in PBS at the same concentration.

110. The formulation of claim 106, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

111. The formulation of claim 106, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

112. The formulation of claim 106, wherein the antibody concentration is at least about 50 mg/mL.

113. An aqueous formulation comprising

mannitol,

polysorbate 80,

an antibody, or antigen-binding fragment thereof, at a concentration of at least about 20 mg/mL, and

water,

wherein the antibody, or antigen-binding fragment, has a hydrodynamic diameter (D_h) of less than about 4 nm, and

wherein the antibody, or antigen-binding fragment thereof, has a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

7, and a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

114. The formulation of claim 113, wherein the antibody, or antigen-binding portion thereof, has a D_h of less than about 3 nm.

115. The formulation of claim 113, wherein the antibody, or antigen-binding portion thereof, has an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

1, and an HCVR comprising the amino acid sequence set forth in SEQ ID NO;

2.

116. The formulation of claim 113, wherein the antibody, or antigen-binding portion thereof, is adalimumab.

117. The formulation of claim 113, wherein the antibody concentration is at least about 50 mg/mL.

120. A method of preparing an aqueous formulation comprising an antibody, or antigen-binding fragment thereof, at a concentration of at least about 20 mg/ml and water, the method comprising:

a) providing the antibody, or antigen-binding fragment thereof, in a first solution; and

b) subjecting the first solution to diafiltration using water as a diafiltration medium until at least five-fold volume exchange with the water has been achieved to thereby prepare the aqueous formulation;

wherein the antibody or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

123. The method of claim 120 or 121, wherein the diafiltration medium consists of water.

124. The method of claim 120 or 121, wherein the first solution is subjected to diafiltration with water until at least about a six-fold volume exchange or at least about a seven-fold volume exchange is achieved.

125. The method of claim 120 or 121, wherein the aqueous formulation has a final concentration of excipients which is at least about 95% less or 99% less than the first solution.

126. The method of claim 120 or 121, wherein the first antibody, or antigen-binding fragment thereof, solution is obtained from a mammalian cell expression system and has been purified to remove host cell proteins (HCPs).

127. The method of claim 120 or 121, wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) selected from the group consisting of greater than about 57 kDa, greater than about 100 kDa, greater than about 150 kDa, greater than about 200 kDa, and greater than about 250 kDa.

128. The method of claim 120 or 121, further comprising adding an excipient to the aqueous formulation.

129. The method of claim 128, wherein the aqueous formulation is a pharmaceutical formulation.

130. The method of claim 120 or 121, wherein the aqueous formulation is a pharmaceutical formulation.

131. The method of claim 130, further comprising loading the aqueous formulation into a device suitable for administering the aqueous formulation to a subject.

132. The method of claim 120 or 121, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of a chimeric antibody, a human antibody, a humanized antibody, and a domain antibody (dAb).

133. The method of claim 120 or 121, wherein the antibody, or antigen-binding fragment thereof, is an anti-TNFα

or an anti-IL-12 antibody.

134. The method of claim 120 or 121, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of, alemtuzumab, arcitumomab, cetuximab, trastuzumab, imciromab pentetate, capromab pendetide, infliximab, abciximab, rituximab, basiliximab, palivizumab, nofetumomab, omalizumab, daclizumab, ibritumomab tiuxetan, muromonab-CD3, edrecolomab gemtuzumab ozogamicin, golimumab, certolizumab pegol, eculizumab, ustekinumab, panitumumab, tositumomab, I¹³¹ tositumomab, and bevacizumab.

135. The formulation of claim 120 or 121, wherein the antibody, or antigen-binding fragment thereof, is adalimumab.

121. A method of preparing an aqueous formulation of an antibody, or antigen-binding fragment thereof, at a concentration of at least about 20 mg/ml, the method comprising:

a) providing the antibody, or an antigen-binding fragment thereof, in a first solution;

b) subjecting the first solution to diafiltration using water as a diafiltration medium until at least a five-fold volume exchange with the water has been achieved to thereby prepare a diafiltered antibody, or antigen-binding fragment thereof, solution; and

c) concentrating the diafiltered antibody, or antigen-binding fragment thereof, solution to thereby prepare the aqueous formulation of the antibody, or antigen-binding fragment thereof;

wherein the antibody, or antigen-binding fragment thereof, has a molecular weight (M_w) greater than about 47 kDa.

122. The method of claim 121, wherein the concentration of the diafiltered antibody, or antigen-binding fragment thereof, solution is achieved via centrifugation.