Claims for Patent: 8,440,799

Summary for Patent: 8,440,799

Title:	Methods of purifying anti A .beta. antibodies
Abstract:	The present application provides methods of purifying A.beta. binding proteins having a Fc region, for example, anti-A.beta. antibodies or antibody fusions, by adsorbing the A.beta. binding protein to a Fc binding agent, such as, for example, Protein A or Protein G, followed by a wash with a divalent cation salt buffer to remove impurities and subsequent recovery of the adsorbed A.beta. binding protein. The present application also features methods of eluting the purified A.beta. binding protein as well as the incorporation of the methods within a purification train. Kits comprising components for carrying out the methods and instructions for use are also provided.
Inventor(s):	Godavarti; Ranganathan (Burlington, MA), Iskra; Timothy (Derry, NH)
Assignee:	Janssen Alzheimer Immunotherapy (Dublin, IE) Wyeth LLC (Madison, NJ)
Application Number:	12/903,053
Patent Claims:	1. A method for purifying an anti-A.beta. antibody having an Fc region from a source liquid comprising the antibody and one or more impurities, comprising the steps of: adsorbing the anti-A.beta. antibody to an Fc binding agent comprising Protein A; washing the Fc binding agent to which the anti-A.beta. antibody is adsorbed with a buffer solution containing CaCl.sub.2 at a concentration of from 0.5 M to 3 M to remove one or more impurities; recovering the anti-A.beta. antibody from the Fc binding agent by eluting the antibody using an elution buffer having a pH in a range from 2 to 4; and subjecting the anti-A.beta. antibody recovered from the Fc binding agent to anion exchange chromatography. 2. The method of claim 1, wherein the anti-A.beta. antibody is selected from the group consisting of: an antibody fusion, a murine antibody, a chimeric antibody, a humanized antibody and a human antibody. 3. The method of claim 1, wherein the anti-A.beta. antibody is a humanized anti-A.beta. antibody. 4. The method of claim 3, wherein the humanized anti-A.beta. antibody is selected from the group consisting of: 3D6, 10D5, 12A11, 266, 15C11, and 12B4. 5. The method of claim 1, wherein the anti-A.beta. antibody is recombinantly produced. 6. The method of claim 5, wherein the anti-A.beta. antibody is recombinantly produced in a Chinese Hamster Ovary (CHO) cell. 7. The method of claim 1, wherein the buffer solution has a pH value in a range between about 4 to about 8. 8. The method of claim 1, wherein the buffer solution has a CaCl.sub.2 concentration in a range between about 0.5 M to about 2 M. 9. The method of claim 1, wherein the steps of adsorbing the anti-A.beta. antibody to an Fc binding agent and washing the Fc binding agent are performed at a temperature in the range between about 2.degree. C. to about 24.degree. C. 10. The method of claim 1, wherein the one or more impurities comprise one or more of a host cell protein, a host cell DNA, a cell culture protein, and mixtures thereof. 11. The method of claim 1, wherein the step of recovering the anti-A.beta. antibody from the Fc binding agent comprises eluting the antibody using an elution buffer having a pH in a range from about 2.5 to about 3.5. 12. The method of claim 1, wherein the method further comprises a chromatography step selected from the group consisting of: cation exchange chromatography, immobilized metal affinity chromatography and hydrophobic interaction chromatography (HIC). 13. The method of claim 1, wherein the method further comprises an additional purification step selected from the group consisting of: hydroxyapatite chromatography, dialysis, affinity chromatography, ammonium sulphate precipitation, ethanol precipitation, reverse phase HPLC (RP-HPLC), and chromatofocusing. 14. The method of claim 4, wherein the humanized anti-A.beta. antibody is a 3D6 antibody comprising a light chain variable region comprising the amino acid sequence of residues 1-112 of SEQ ID NO:1 and a heavy chain variable region comprising the amino acid sequence of residues 1-119 of SEQ ID NO:2. 15. The method of claim 14, wherein the humanized anti-A.beta. antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:1 and a heavy chain comprising the amino acid sequence of residues 1-448 of SEQ ID NO:2. 16. The method of claim 1, wherein the anti-A.beta. antibody is of isotype IgM, IgG1, IgG2, IgG3, or IgG4. 17. The method of claim 16, wherein the antibody is of isotype human IgG1. 18. The method of claim 1, wherein the one or more impurities comprise one or more of an intron read through variant species, an under-disulfide bonded species, or a low-molecular weight species. 19. The method of claim 1, wherein the anion exchange chromatography step is performed using an anion exchange resin comprising TMAE. 20. The method of claim 9, wherein the temperature is in a range of from 18.degree. C. to 24.degree. C. 21. The method of claim 1, further comprising filtering the anti-A.beta. antibody after the anion exchange chromatography, wherein the filtering comprises one or more of a virus filtration step, an ultrafiltration/diafiltration step, or a microbial contaminant filtration step. 22. The method of claim 21, wherein the anti-A.beta. antibody comprises a light chain variable region comprising the amino acid sequence of residues 1-112 of SEQ ID NO:1 and a heavy chain variable region comprising the amino acid sequence of residues 1-119 of SEQ ID NO:2. 23. The method of claim 22, wherein the anti-A.beta. antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:1 and a heavy chain comprising the amino acid sequence of residues 1-448 of SEQ ID NO:2.

Details for Patent 8,440,799

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2025-06-17
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2025-06-17
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2025-06-17