ANTIBODY FORMULATIONS AND METHODS

1 . A pharmaceutical formulation comprising:

(a) a chimeric or humanized version of antibody 2A4 (ATCC Accession Number PTA-9662) or of antibody 7D8 (ATCC Accession Number PTA-9468), or fragment thereof, that specifically competes for binding to antigen with 2A4 or 7D8, wherein the antibody is present at a concentration within the range from about 1 mg/mL to about 100 mg/mL;

(b) histidine buffer present at a concentration within the range from about 20 mM to about 30 mM;

(c) trehalose present at a concentration within the range from about 210 mM to about 250 mM; and

(d) polysorbate 20 present at a concentration within the range from about 0.005% to about 0.05% by weight;

wherein the formulation is characterized by a pH within the range from about 6 to about 7.

2 . The formulation of claim 1 , wherein the antibody is a humanized version of antibody 2A4.

3 . The formulation of claim 1 , wherein the antibody comprises a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 4.

4 . The formulation of claim 1 , wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 5.

5 . The formulation of claim 3 , wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 5.

6 . The formulation of claim 5 , wherein the antibody comprises a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as any one of SEQ ID NO: 14-16.

7 . The formulation of claim 6 , wherein the antibody comprises a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 15.

8 . The formulation of claim 1 , wherein the antibody comprises a light chain variable region comprising three complementarity determining regions set forth as SEQ ID NOs: 6, 7, and 8, and a heavy chain variable region comprising three complementarity regions set forth as SEQ ID NOs: 9, 10, and 11.

9 . The formulation of claim 1 , wherein the antibody is a humanized or chimeric version of antibody 7D8 produced by ATCC Accession Number PTA-9468.

10 . The formulation of claim 1 , wherein the antibody comprises a light chain variable region comprising three complementarity determining regions set forth as SEQ ID NOs: 12, 7, and 8, and a heavy chain variable region comprising three complementarity regions set forth as SEQ ID NOs: 9, 10, and 11.

11 . The formulation of any one of claims 1 - 10 , wherein the antibody is present at a concentration within the range from about 5 mg/mL to about 15 mg/mL.

12 . The formulation of claim 11 , wherein the antibody is present at a concentration of about 10 mg/mL.

13 . The formulation of any one of claims 1 - 10 , wherein the antibody is present at a concentration within the range from about 25-75 mg/mL.

14 . The formulation of claim 13 , wherein the antibody is present at a concentration of about 50 mg/mL.

15 . The formulation of any one of claims 1 - 14 , wherein the histidine buffer is present at a concentration of about 25 mM.

16 . The formulation of claim 15 , wherein the histidine buffer comprises L-histidine and L-histidine HCl monohydrate.

17 . The formulation of claim 16 , wherein the L-histidine is present at a concentration within the range from about 16 mM to about 22 mM and the L-histidine HCl monohydrate is present at a concentration within the range from about 4 mM to about 8 mM.

18 . The formulation of any one of claims 1 - 17 , wherein the trehalose is present at a concentration of about 230 mM.

19 . The formulation of any one of claims 1 - 18 , which is characterized by an osmolality of about 300 mOsm/kg.

20 . The formulation of any one of claims 1 - 19 , wherein less than about 10% of the antibody is present as an aggregate in the formulation.

21 . The formulation of any one of claims 1 - 20 , which further comprises a bulking agent.

22 . The formulation of any one of claims 1 - 21 , which is sterile.

23 . The formulation of any one of claims 1 - 22 , which is stable upon freezing and thawing.

24 . A pharmaceutical formulation comprising:

(a) an antibody comprising a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as any one of SEQ ID NOs: 14-16, and which is present at a concentration of about 10 mg/mL;

(b) a histidine buffer present at a concentration of about 25 mM;

(c) trehalose present at a concentration of about 230 mM;

(d) polysorbate 20 present at a concentration of about 0.2 g/L; and

(e) a pH of about 6.5.

25 . The formulation of claim 24 , wherein the antibody comprises a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 15.

26 . A pharmaceutical formulation comprising:

(a) an antibody, which is antibody 2A4 (ATCC Accession Number PTA-9662), antibody 7D8 (ATCC Accession Number PTA-9468), or a chimeric or humanized version of antibody 2A4 or of antibody 7D8, or fragment thereof, that specifically competes for binding to antigen with 2A4 or 7D8, wherein the antibody is present at a concentration within the range from about 50 mg/mL to about 100 mg/mL;

(b) a buffer;

(c) a non-reducing sugar; and

(d) a non-ionic surfactant.

27 . A lyophilized formulation of an antibody, comprising

(a) a humanized version of antibody 2A4 (ATCC Accession Number PTA-9662) or antibody 7D8 (ATCC Accession Number PTA-9468) or antigen binding fragment thereof;

(b) histidine;

(c) trehalose; and

(d) polysorbate 20.

28 . The lyophilized formulation of claim 27 , wherein the formulation has a pH of between about 6 to about 7 when reconstituted.

29 . The lyophilized formulation of claim 28 , wherein the formulation has a pH of about 6.5 when reconstituted.

30 . The lyophilized formulation of claim 27 , comprising about 100 mg to about 1000 mg of the antibody.

31 . The lyophilized formulation of claim 27 , wherein the polysorbate 20 is present at a concentration within the range from about 0.005% to about 0.05% by weight.

32 . The lyophilized formulation of claim 27 , that enables reconstitution to yield an aqueous solution comprising:

(a) an antibody comprising a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as any one of SEQ ID NOs: 14-16, and which is present at a concentration of about 10 mg/mL;

(b) a histidine buffer present at a concentration of about 25 mM;

(c) trehalose present at a concentration of about 230 mM;

(d) polysorbate 20 present at a concentration of about 0.2 g/L; and

(e) a pH of about 6.5.

33 . The lyophilized formulation of claim 32 , wherein the lyophilized formulation comprises about 100 mg of the antibody and enables reconstitution with sterile water.

34 . A nucleic acid comprising a nucleotide sequence encoding SEQ ID NO: 13.

35 . The nucleic acid of claim 34 comprising the nucleotide sequence of SEQ ID NO: 19.

36 . The nucleic acid of claim 35 comprising the nucleotide sequence of SEQ ID NO: 20.

37 . A nucleic acid comprising a nucleotide sequence encoding any one of SEQ ID NOs: 14-16.

38 . The nucleic acid of claim 37 comprising a nucleotide sequence encoding SEQ ID NO: 15.

39 . The nucleic acid of claim 38 comprising the nucleotide sequence of SEQ ID NO: 22.

40 . The nucleic acid of claim 39 comprising the nucleotide sequence of SEQ ID NO: 23.

41 . A vector comprising the nucleic acid of any one of claims 34 - 40 .

42 . A vector comprising the nucleic acids of claims 34 and 38 .

43 . A vector comprising the nucleic acids of claims 35 and 39 .

44 . A vector comprising the nucleic acids of claims 36 and 40 .

45 . A host cell having stably incorporated into its genome the nucleic acid of any of claims 34 - 40 .

46 . The host cell of claim 45 , wherein the host cell is a CHO cell.

47 . The host cell of claim 45 having stably incorporated into its genome a nucleic acid comprising the nucleotide sequence encoding SEQ ID NO: 13 and a nucleic acid comprising the nucleotide sequence encoding any one of SEQ ID NOs: 14-16.

48 . The host cell of claim 38 having stably incorporated into its genome a nucleic acid comprising the nucleotide sequence encoding SEQ ID NO: 19 and a nucleic acid comprising the nucleotide sequence encoding SEQ ID NO: 22.

49 . The host cell of claim 38 having stably incorporated into its genome a nucleic acid comprising the nucleotide sequence encoding SEQ ID NO: 20 and a nucleic acid comprising the nucleotide sequence encoding SEQ ID NO: 23.

50 . The host cell of any one of claims 47 - 49 , wherein the host cell is a CHO cell.

51 . A method of making a pharmaceutical formulation comprising:

(a) culturing mammalian cells having stably incorporated into their genome one or more nucleic acids encoding the light and heavy chains of a humanized 2A4 antibody so that the cells secrete the antibody into the cell culture media, and purifying the antibody from the cell culture media;

(b) and preparing the formulation of claim 1 .

52 . The method of claim 51 , wherein the nucleic acid encoding the light chain of a humanized 2A4 antibody comprises a nucleotide sequence encoding SEQ ID NO: 13, and wherein the nucleic acid encoding the heavy chain of a humanized 2A4 antibody comprises a nucleotide sequence encoding any one of SEQ ID NOs: 14-16.

53 . The method of claim 52 , wherein the nucleic acid encoding the light chain of a humanized 2A4 antibody comprises a nucleotide sequence encoding SEQ ID NO: 13, and wherein the nucleic acid encoding the heavy chain of a humanized 2A4 antibody comprises a nucleotide sequence encoding SEQ ID NO: 15.

54 . The method of claim 53 , wherein the nucleic acid encoding the light chain of a humanized 2A4 antibody comprises the nucleotide sequence of SEQ ID NO: 19, and wherein the nucleic acid encoding the heavy chain of a humanized 2A4 antibody comprises the nucleotide sequence of SEQ ID NO: 22.

55 . The method of claim 54 , wherein the nucleic acid encoding the light chain of a humanized 2A4 antibody comprises the nucleotide sequence of SEQ ID NO: 20, and wherein the nucleic acid encoding the heavy chain of a humanized 2A4 antibody comprises the nucleotide sequence of SEQ ID NO: 23.

56 . The method of any one of claims 51 - 55 , further comprising evaluating at least one property of antibody in the formulation selected from the group consisting of the physical stability, chemical stability and biological activity.

57 . The method of any one of claims 51 - 55 , wherein the mammalian cells are CHO cells.

58 . A method of therapeutically or prophylactically treating a human patient having or at risk for having amyloidosis characterized by the presence of amyloid protein fibrils, the method comprising administering to the patient an effective dosage of the formulation of any one of claims 1 - 33 .

59 . The method of claim 58 , wherein the human patient has amyloid A amyloidosis characterized by the presence of amyloid A protein fibrils.

60 . The method of claim 59 , wherein the formulation is the formulation of claim 25 .

61 . The method of claim 60 , wherein the human patient has AL amyloidosis characterized by the presence of amyloid light chain-type protein fibrils.

62 . The method of claim 61 , wherein the formulation is the formulation of claim 25 .

63 . The method of claim 61 or 62 , wherein the patient is also treated with one or both of melphalan, bortezomib, lenolidomide or carfilzomib.

64 . The method of claim 63 , wherein the patient is treated with melphalan and/or bortezomib prior to treatment with the formulation of any one of claims 1 - 33 .

65 . The method of claim 64 , wherein the patient is treated with melphalan and/or bortezomib concurrently with treatment with the formulation of any one of claims 1 - 33 .

66 . The method of claim 64 , wherein the patient is treated with melphalan and/or bortezomib subsequent to treatment with the formulation of any one of claims 1 - 33 .

67 . The method of any one of claims 58 - 66 , wherein the AL amyloidosis is associated with a dyscrasia of the B lymphocyte lineage.

68 . The method of claim 67 , wherein the dyscrasia is a malignancy.

69 . The method of claim 68 , wherein the malignancy is multiple myeloma.

70 . The method of any one of claims 58 - 69 , wherein the formulation is administered in multiple dosages.

71 . The method of claim 70 , wherein the formulation is administered at a frequency in a range of about daily to about annually.

72 . The method of claim 71 , wherein the frequency is in a range of about every other week to about every three months.

73 . The method of any one of claims 58 - 72 , wherein the formulation is administered intravenously at a dose in a range from about 10 mg to about 5000 mg humanized 2A4 drug substance.

74 . The method of claim 73 , wherein the formulation is administered at a dose in a range from about 30 mg to about 2500 mg humanized 2A4 drug substance at a frequency in a range of about every other week to about every other month.

75 . The method of claim 73 or 74 , wherein the formulation is administered once a month.

76 . The method of any one of claims 73 - 75 , wherein the dose is about 30 mg humanized 2A4 drug substance.

77 . The method of any one of claims 73 - 75 , wherein the dose is about 100 mg humanized 2A4 drug substance.

78 . The method of any one of claims 73 - 75 , wherein the dose is about 300 mg humanized 2A4 drug substance.

79 . The method of any one of claims 73 - 75 , wherein the dose is about 1000 mg humanized 2A4 drug substance.

80 . The method of any one of claims 73 - 75 , wherein the dose is about 2500 mg humanized 2A4 drug substance.

81 . A method of therapeutically or prophylactically treating a human patient having or at risk for having light chain (AL) amyloidosis characterized by the presence of amyloid fibrils, deposits or prefibrillar aggregates, comprising administering to the patient an effective dosage of a pharmaceutical formulation comprising:

(a) an antibody comprising a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as any one of SEQ ID NOs: 14-16, and which is present at a concentration of about 10 mg/mL;

(b) a histidine buffer present at a concentration of about 25 mM;

(c) trehalose present at a concentration of about 230 mM;

(d) polysorbate 20 present at a concentration of about 0.2 g/L; and

(e) a pH of about 6.5.

82 . The method of claim 81 , wherein the dosage is from about 0.5 mg/kg to about 30 mg/kg of the antibody administered intravenously or subcutaneously at a frequency of from about weekly to about quarterly.

83 . The method of claim 82 , wherein the frequency of administration is once every 28 days.

84 . The method of claim 82 , wherein the dosage is about 0.5 mg/kg to about 8 mg/kg.

85 . The method of claim 82 , wherein the dosage is about 8 mg/kg to about 30 mg/kg.

86 . A pharmaceutical product, comprising:

(a) a vial comprising about 100 mg antibody in powder form;

(b) instructions for reconstitution of the antibody; and

(c) instructions for preparing the reconstituted antibody for infusion,

wherein:

(i) the antibody comprises a light chain comprising an amino acid sequence set forth as SEQ ID NO: 13 and a heavy chain comprising an amino acid sequence set forth as any one of SEQ ID NOs: 14-16; and

(ii) the reconstitution instructions require reconstitution with water for injection to an extractable volume of 10 mL.