Methods of purifying anti-interleukin-13 antibodies
The invention relates to a pharmaceutical composition comprising an interleukin-13 antibody, more particularly a monoclonal interleukin-13 antibody, especially a human interleukin-13 monoclonal antibody, to a process for purifying said antibody and to the use of said composition in treating interleukin-13 related disorders, such as asthma, atopic dermatitis, allergic rhinitis, fibrosis, chronic obstructive pulmonary disease, scleroderma, inflammatory bowel disease and Hodgkin's lymphoma, particularly asthma.
1. A process for purifying an IL-13 antibody comprising more than one chromatographic separation steps, wherein each of said separation steps comprises elution with a single acetate buffer comprising 50 mM sodium acetate and 85 mM sodium chloride buffered to a pH of 3.5-7.0.
2. The process of claim 1 , wherein the chromatographic separation steps are selected from the group consisting of affinity chromatography and ion exchange chromatography.
3. The process of claim 2 , wherein the chromatographic separation is performed by Protein A affinity chromatography followed by cation exchange chromatography followed by anion exchange chromatography.
4. The process of any one of claims 1 to 3 , wherein the elution buffer is buffered to pH 5.5±0.1.
5. The process of any one of claims 1 to 3 , wherein the IL-13 antibody is a human monoclonal antibody.
6. The process of any one of claims 1 to 3 , wherein the IL-13 antibody is selected from the group consisting of:
(a) An antibody comprising a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:9, HCDR2 has the amino acid sequence of SEQ ID NO:10, HCDR3 has the amino acid sequence of SEQ ID NO:11, LCDR1 has the amino acid sequence of SEQ ID NO:12, LCDR2 has the amino acid sequence of SEQ ID NO:13, and LCDR3 has the amino acid sequence of SEQ ID NO:14;
(b) An antibody comprising a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:3, HCDR2 has the amino acid sequence of SEQ ID NO:4, HCDR3 has the amino acid sequence of SEQ ID NO:5, LCDR1 has the amino acid sequence of SEQ ID NO:6, LCDR2 has the amino acid sequence of SEQ ID NO:7, and LCDR3 has the amino acid sequence of SEQ ID NO:8;
(c) An antibody comprising a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:15, HCDR2 has the amino acid sequence of SEQ ID NO:16, HCDR3 has the amino acid sequence of SEQ ID NO:17, LCDR1 has the amino acid sequence of SEQ ID NO:18, LCDR2 has the amino acid sequence of SEQ ID NO:19, and LCDR3 has the amino acid sequence of SEQ ID NO:20;
(d) An antibody comprising a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:21, HCDR2 has the amino acid sequence of SEQ ID NO:22, HCDR3 has the amino acid sequence of SEQ ID NO:23, LCDR1 has the amino acid sequence of SEQ ID NO:24, LCDR2 has the amino acid sequence of SEQ ID NO:25, and LCDR3 has the amino acid sequence of SEQ ID NO:26;
(e) An antibody comprising a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:27, HCDR2 has the amino acid sequence of SEQ ID NO:28, HCDR3 has the amino acid sequence of SEQ ID NO:29, LCDR1 has the amino acid sequence of SEQ ID NO:30, LCDR2 has the amino acid sequence of SEQ ID NO:31, and LCDR3 has the amino acid sequence of SEQ ID NO:32; and
(f) An antibody comprising a heavy chain variable region (VH) having the amino acid sequence of SEQ ID NO:1, and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO:2.
7. The process of claim 6 , wherein the IL-13 antibody comprises a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:3, HCDR2 has the amino acid sequence of SEQ ID NO:4, HCDR3 has the amino acid sequence of SEQ ID NO:5, LCDR1 has the amino acid sequence of SEQ ID NO:6, LCDR2 has the amino acid sequence of SEQ ID NO:7, and LCDR3 has the amino acid sequence of SEQ ID NO:8.
8. The process of claim 6 , wherein the IL-13 antibody comprises a heavy chain variable region (VH) having the amino acid sequence of SEQ ID NO:1, and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO:2.
9. The process of claim 6 , wherein the IL-13 antibody is present after elution in an amount of about 50 mg/ml, about 100 mg/ml or about 150 mg/ml.
10. The process of claim 9 , wherein the IL-13 antibody is present after elution in an amount of about 150 mg/ml.
11. The process of claim 4 , wherein the IL-13 antibody comprises a set of complementarity determining regions (CDRs) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein HCDR1 has the amino acid sequence of SEQ ID NO:3, HCDR2 has the amino acid sequence of SEQ ID NO:4, HCDR3 has the amino acid sequence of SEQ ID NO:5, LCDR1 has the amino acid sequence of SEQ ID NO:6, LCDR2 has the amino acid sequence of SEQ ID NO:7, and LCDR3 has the amino acid sequence of SEQ ID NO:8.
12. The process of claim 11 , wherein the IL-13 antibody comprises a heavy chain variable region (VH) having the amino acid sequence of SEQ ID NO:1, and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO:2.
13. The process of claim 11 , wherein the IL-13 antibody is present after elution in an amount of about 50 mg/ml, about 100 mg/ml or about 150 mg/ml.
14. The process of claim 11 , wherein the IL-13 antibody is present after elution in an amount of about 150 mg/ml.