Human antibodies that bind human il-12 and methods for producing

Abstract

Human antibodies, preferably recombinant human antibodies, that specifically bind to human interleukin-12 (hIL-12) are disclosed. Preferred antibodies have high affinity for hIL-12 and neutralize hIL-12 activity in vitro and in vivo. An antibody of the invention can be a full-length antibody or an antigen-binding portion thereof. The antibodies, or antibody portions, of the invention are useful for detecting hIL-12 and for inhibiting hIL-12 activity, e.g., in a human subject suffering from a disorder in which hIL-12 activity is detrimental. Nucleic acids, vectors and host cells for expressing the recombinant human antibodies of the invention, and methods of synthesizing the recombinant human antibodies, are also encompassed by the invention.

1 . A method of treating a human subject suffering from an autoimmune disorder, comprising administering to the human subject an effective amount of a human antibody, or antigen-binding portion thereof, which binds to an epitope of the p40 subunit of IL-12, thereby treating said subject. 
     
     
         2 . The method of  claim 1 , wherein the autoimmune disorder is psoriasis. 
     
     
         3 . The method of  claim 1 , wherein the autoimmune disorder is rheumatoid arthritis. 
     
     
         4 . The method of  claim 1 , wherein the autoimmune disorder is Crohn's disease. 
     
     
         5 . The method of  claim 1 , wherein the autoimmune disorder is Multiple Sclerosis. 
     
     
         6 . The method of  claim 1 , wherein the antibody, or antigen-binding portion thereof, binds to the epitope of the p40 subunit when the p40 subunit is bound to the p35 subunit of IL-12. 
     
     
         7 . The method of  claim 1 , wherein the antibody, or antigen-binding portion thereof, binds to the epitope of the p40 subunit when the p40 subunit is bound to a p19 subunit. 
     
     
         8 . The method of  claim 1 , wherein the antibody, or antigen-binding portion thereof, binds to the epitope of the p40 subunit when the p40 subunit is bound to the p35 subunit of IL-12 and when the p40 subunit is bound to a p19 subunit. 
     
     
         9 . The method of  claim 1 , wherein the antibody, or antigen binding portion thereof, binds to an epitope of the p40 subunit of IL-12 to which an antibody selected from the group consisting of Y61 and J695 binds. 
     
     
         10 . The method of  claim 1 , wherein the antibody further binds to a first heterodimer and also binds to a second heterodimer, wherein the first heterodimer comprises the p40 subunit of IL-12 and the p35 subunit of IL-12, and wherein the second heterodimer comprises the p40 subunit of IL-12 and a p19 subunit. 
     
     
         11 . The method of  claim 10 , wherein the antibody neutralizes the biological activity of the first heterodimer. 
     
     
         12 . The method of  claim 10 , wherein the antibody neutralizes the biological activity of the second heterodimer. 
     
     
         13 . The method of  claim 10 , wherein the antibody neutralizes the biological activity of the first heterodimer and the second heterodimer. 
     
     
         14 . The method of  claim 10 , wherein the antibody, or antigen binding portion thereof, inhibits phytohemagglutinin blast proliferation in an in vitro PHA assay with an IC 50  of 1×10 −9  M or less, or which inhibits human IFNγ production with an IC 50  of 1.6×10 −10  M or less. 
     
     
         15 . The method of  claim 1 , wherein the antibody, or antigen binding portion thereof, dissociates from the p40 subunit of IL-12 with a K d  of 1.34×10 −10  M or less or a k off  rate constant of 1×10 −3  s −1  or less, as determined by surface plasmon resonance. 
     
     
         16 . The method of  claim 1 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 25 and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 26; 
     
     
         17 . The method of  claim 16 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27 and a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 28. 
     
     
         18 . The method of  claim 16 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29 and a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30. 
     
     
         19 . A method of treating a human subject suffering from an autoimmune disorder, comprising administering to the human subject an effective amount of a human antibody, or antigen-binding portion thereof, which binds to an interleukin comprising a p40 subunit, thereby treating said subject. 
     
     
         20 . The method of  claim 19 , wherein the autoimmune disorder is psoriasis. 
     
     
         21 . The method of  claim 19 , wherein the autoimmune disorder is rheumatoid arthritis. 
     
     
         22 . The method of  claim 19 , wherein the autoimmune disorder is Crohn's disease. 
     
     
         23 . The method of  claim 19 , wherein the autoimmune disorder is Multiple Sclerosis. 
     
     
         24 . The method of  claim 19 , wherein the interleukin comprises a p40 subunit and a p35 subunit. 
     
     
         25 . The method of  claim 24 , wherein the interleukin is IL-12. 
     
     
         26 . The method of  claim 19 , wherein the interleukin comprises a p40 subunit and a p19 subunit. 
     
     
         27 . The method of  claim 19 , wherein the antibody, or antigen binding portion thereof, binds to an epitope of the p40 subunit. 
     
     
         28 . The method of  claim 19 , wherein the antibody, or antigen binding portion thereof, binds to an epitope of the p40 subunit to which an antibody selected from the group consisting of Y61 and J695 binds. 
     
     
         29 . The method of  claim 19 , wherein the antibody, or antigen binding portion thereof, dissociates from the p40 subunit of the interleukin with a K d  of 1.34×10 −10  M or less or a k off  rate constant of 1×10 −3 s −1  or less, as determined by surface plasmon resonance. 
     
     
         30 . The method of  claim 19 , wherein the antibody, or antigen binding portion thereof, neutralizes the biological activity of the interleukin. 
     
     
         31 . The method of  claim 30 , wherein the antibody, or antigen binding portion thereof, inhibits phytohemagglutinin blast proliferation in an in vitro PHA assay with an IC 50  of 1×10 −9  M or less, or which inhibits human IFNγ production with an IC 50  of 1.6×10 −10  M or less. 
     
     
         32 . The method of  claim 19 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 25 and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 26. 
     
     
         33 . The method of  claim 32 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27 and a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 28. 
     
     
         34 . The method of  claim 32 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29 and a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30. 
     
     
         35 . A method of treating a human subject suffering from psoriasis, comprising administering to the human subject an effective amount of a human antibody, or antigen-binding portion thereof, which binds to an epitope of the p40 subunit of IL-12, thereby treating the subject. 
     
     
         36 . The method of  claim 35 , wherein the subject exhibits an improvement in skin condition for an extended period following administration of the antibody, or antigen-binding portion thereof. 
     
     
         37 . The method of  claim 36 , wherein the subject exhibits flattening of plaques. 
     
     
         38 . The method of  claim 36 , wherein the subject exhibits a decrease in scaling. 
     
     
         39 . The method of  claim 35 , wherein the subject exhibits a total clearance of plaques for an extended period following administration of the antibody, or antigen-binding portion thereof. 
     
     
         40 . The method of  claim 35 , wherein said effective amount is about 0.01 mg/kg to about 10 mg/kg. 
     
     
         41 . The method of  claim 35 , wherein said antibody, or antigen-binding portion thereof, is administered to said subject subcutaneously. 
     
     
         42 . The method of  claim 35 , wherein said antibody, or antigen-binding portion thereof, is administered to said subject intravenously. 
     
     
         43 . The method of  claim 35 , wherein the antibody, or antigen-binding portion thereof, binds to the epitope of the p40 subunit when the p40 subunit is bound to the p35 subunit of IL-12. 
     
     
         44 . The method of  claim 35 , wherein the antibody, or antigen-binding portion thereof, binds to the epitope of the p40 subunit when the p40 subunit is bound to a p19 subunit. 
     
     
         45 . The method of  claim 35 , wherein the antibody, or antigen-binding portion thereof, binds to the epitope of the p40 subunit when the p40 subunit is bound to the p35 subunit of IL-12 and when the p40 subunit is bound to a p19 subunit. 
     
     
         46 . The method of  claim 35 , wherein the antibody, or antigen binding portion thereof, binds to an epitope of the p40 subunit of IL-12 to which an antibody selected from the group consisting of Y61 and J695 binds. 
     
     
         47 . The method of  claim 35 , wherein the antibody further binds to a first heterodimer and also binds to a second heterodimer, wherein the first heterodimer comprises the p40 subunit of IL-12 and the p35 subunit of IL-12, and wherein the second heterodimer comprises the p40 subunit of IL-12 and a p19 subunit. 
     
     
         48 . The method of  claim 47 , wherein the antibody neutralizes the biological activity of the first heterodimer. 
     
     
         49 . The method of  claim 47 , wherein the antibody neutralizes the biological activity of the second heterodimer. 
     
     
         50 . The method of  claim 47 , wherein the antibody neutralizes the biological activity of the first heterodimer and the second heterodimer. 
     
     
         51 . The method of  claim 47 , wherein the antibody, or antigen binding portion thereof, inhibits phytohemagglutinin blast proliferation in an in vitro PHA assay with an IC 50  of 1×10 −9  M or less, or which inhibits human IFNγ production with an IC 50  of 1.6×10 −10  M or less. 
     
     
         52 . The method of  claim 35 , wherein the antibody, or antigen binding portion thereof, dissociates from the p40 subunit of IL-12 with a K d  of 1.34×10 −10  M or less or a k off  rate constant of 1×10 −3 s −1  or less, as determined by surface plasmon resonance. 
     
     
         53 . The method of  claim 35 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 25 and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 26. 
     
     
         54 . The method of  claim 53 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27 and a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 28. 
     
     
         55 . The method of  claim 53 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29 and a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30. 
     
     
         56 . A method for inhibiting the activity of an interleukin comprising a p40 subunit in a human subject suffering from psoriasis, comprising administering to the human subject an effective amount of a human antibody, or antigen-binding portion thereof, which to an interleukin comprising a p40 subunit, such that the psoriasis is treated. 
     
     
         57 . The method of  claim 56 , wherein the subject exhibits an improvement in skin condition for an extended period following administration of the antibody, or antigen-binding portion thereof. 
     
     
         58 . The method of  claim 57 , wherein the subject exhibits flattening of plaques. 
     
     
         59 . The method of  claim 57 , wherein the subject exhibits a decrease in scaling. 
     
     
         60 . The method of  claim 56 , wherein the subject exhibits a total clearance of plaques for an extended period following administration of the antibody, or antigen-binding portion thereof. 
     
     
         61 . The method of  claim 56 , wherein said effective amount is about 0.01 mg/kg to about 10 mg/kg. 
     
     
         62 . The method of  claim 56 , wherein said antibody, or antigen-binding portion thereof, is administered to said subject subcutaneously. 
     
     
         63 . The method of  claim 56 , wherein said antibody, or antigen-binding portion thereof, is administered to said subject intravenously. 
     
     
         64 . The method of  claim 56 , wherein the interleukin comprises a p40 subunit and a p35 subunit. 
     
     
         65 . The method of  claim 64 , wherein the interleukin is IL-12. 
     
     
         66 . The method of  claim 56 , wherein the interleukin comprises a p40 subunit and a p19 subunit. 
     
     
         67 . The method of  claim 56 , wherein the antibody, or antigen binding portion thereof, binds to an epitope of the p40 subunit. 
     
     
         68 . The method of  claim 56 , wherein the antibody, or antigen binding portion thereof, binds to an epitope of the p40 subunit to which an antibody selected from the group consisting of Y61 and J695 binds. 
     
     
         69 . The method of  claim 56 , wherein the antibody, or antigen binding portion thereof, dissociates from the p40 subunit of the interleukin with a K d  of 1.34×10 −10  M or less or a k off  rate constant of 1×10 −3 s −1  or less, as determined by surface plasmon resonance. 
     
     
         70 . The method of  claim 56 , wherein the antibody, or antigen binding portion thereof, neutralizes the biological activity of the interleukin. 
     
     
         71 . The method of  claim 70 , wherein the antibody, or antigen binding portion thereof, inhibits phytohemagglutinin blast proliferation in an in vitro PHA assay with an IC 50  of 1×10 −9  M or less, or which inhibits human IFNγ production with an IC 50  of 1.6×10 −10  M or less. 
     
     
         72 . The method of  claim 56 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 25 and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 26; 
     
     
         73 . The method of  claim 72 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27 and a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 28. 
     
     
         74 . The method of  claim 72 , wherein the antibody, or antigen binding portion thereof, comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 29 and a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 30. 
     
     
         75 . A method of treating a human subject suffering from psoriasis, comprising administering to the human subject about 0.01 mg/kg to about 10 mg/kg of a human antibody, or antigen-binding portion thereof, which binds to an epitope of the p40 subunit of IL-12, thereby treating said subject. 
     
     
         76 . The method of  claim 75 , wherein said antibody, or antigen-binding portion thereof, is administered to said subject subcutaneously. 
     
     
         77 . The method of  claim 75 , wherein said antibody, or antigen-binding portion thereof, is administered to said subject intravenously. 
     
     
         78 . The method of  claim 75 , wherein said antibody, or antigen-binding portion thereof, is administered in an amount of about 1 mg/kg to about 10 mg/kg. 
     
     
         79 . The method of  claim 75 , wherein said antibody, or antigen-binding portion thereof, is administered using a formulation further comprising at least one additional agent selected from the group consisting of a buffer, a polyalcohol and a surfactant. 
     
     
         80 . The method of  claim 79 , wherein said buffer is selected from the group consisting of L-histidine, sodium succinate, sodium citrate, sodium phosphate and potassium phosphate. 
     
     
         81 . The method of  claim 79 , wherein said polyalcohol is selected from the group consisting of mannitol and sorbitol. 
     
     
         82 . The method of  claim 79 , wherein said surfactant is selected from the group consisting of polysorbate 80, polysorbate 20 and BRIJ surfactants. 
     
     
         83 . The method of  claim 79 , wherein said formulation further comprises methionine. 
     
     
         84 . The method of  claim 79 , wherein said formulation comprises a polyalcohol, a surfactant, and a buffer comprising L-histidine. 
     
     
         85 . The method of  claim 84 , wherein the polyalcohol is mannitol. 
     
     
         86 . The method of  claim 84 , wherein the surfactant is polysorbate 80. 
     
     
         87 . The method of  claim 84 , wherein said formulation further comprises methionine. 
     
     
         88 . The method of  claim 87 , wherein said composition comprises
 (a) 1-10% mannitol,   (b) 0-0.05% polysorbate-80,   (c) 1-50 mM methionine, and   (d) a buffer comprising 1-50 mM L-histidine, with a pH of 5 to 7.   
     
     
         89 . The method of  claim 87 , wherein said composition comprises
 (a) 2-4% mannitol,   (b) 0.005-0.01% polysorbate-80,   (c) 5-10 mM methionine, and   (d) a buffer comprising 5-10 mM L-histidine, with a pH of 5 to 7.   
     
     
         90 . A method of treating a subject suffering from psoriasis, comprising subcutaneously administering to the subject a human antibody, or antigen-binding portion thereof, which binds to an epitope of the p40 subunit of IL-12, wherein said antibody, or antigen-binding portion thereof, is administered using a formulation comprising:
 (a) about 1-10 mg/kg of said antibody, or antigen-binding portion thereof,   (b) 2-4% mannitol,   (c) 0.005-0.01% polysorbate-80,   (d) 5-10 mM methionine, and   (e) a buffer comprising 5-10 mM L-histidine, with a pH of 5 to 7, thereby treating said subject.