Protein formulation

Abstract

The present invention relates to methods of minimizing the aggregation of BiTE® molecules in solution. In one embodiment, a method of the invention is used to minimize the aggregation of BiTE® molecules in solution. In another embodiment, a method of the invention is used to minimize the aggregation of BiTE® molecules comprising a first and a second binding site specific for the CD3 and CD19 antigens, respectively, wherein said molecule is in solution. In a specific embodiment, the present invention provides methods to minimize the aggregation of the MT103™ BiTE® molecule in solution.

1 - 76 . (canceled) 
     
     
         77 . A method of preventing, managing, treating or ameliorating B cell disorders or diseases in a human subject in need thereof, said method comprising administering to the subject a prophylactically or therapeutically effective amount of a liquid pharmaceutical formulation comprising a bispecific antibody, wherein the bispecific antibody comprises a first binding domain that specifically binds the CD3 T cell surface antigen and a second binding domain that specifically binds an antigen selected from CD19, CD20, CD22, EphA2, EphA4, INFR, ICOS, Ep-CAM, CEA, and IL-5 receptor. 
     
     
         78 . The method of  claim 77 , wherein the B cell disorder or disease is selected from B cell malignancies, autoimmune diseases, graft-versus-host disease (GVHD), humoral rejection, and post-transplantation lymphoproliferative disorder. 
     
     
         79 . The method of  claim 77 , wherein the B cell disorder includes B cell malignancies. 
     
     
         80 . The method of  claim 79 , wherein the B cell malignancy is selected from B cell subtype non-Hodgkin's lymphoma (NHL); Burkitt's lymphoma; multiple myeloma; pre-B acute lymphoblastic leukemia; malignancies that derive from early B cell precursors; common acute lymphocytic leukemia (ALL); chronic lymphocytic leukemia (CLL); hairy cell leukemia; Null-acute lymphoblastic leukemia; Waldenstrom's Macroglobulinemia; diffuse large B cell lymphoma (DLBCL); pro-lymphocytic leukemia; light chain disease; plasmacytoma; osteosclerotic myeloma; plasma cell leukemia; monoclonal gammopathy of undetermined significance (MGUS); smoldering multiple myeloma (SMM); indolent multiple myeloma (IMM); Hodgkin's lymphoma including classical and nodular lymphocyte pre-dominant type; lymphoplasmacytic lymphoma (LPL); and marginal-zone lymphoma including gastric mucosal-associated lymphoid tissue (MALT) lymphoma. 
     
     
         81 . The method of  claim 80 , wherein the B cell malignancy is selected from:
 (a) non-Hodgkin's lymphoma selected from low grade/follicular NHL, small lymphocytic (SL) NHL, intermediate grade/follicular NHL, intermediate grade diffuse NHL, high grade immunoblastic NHL, high grade lymphoblastic NHL, high grade small non-cleaved cell NHL; mantle-cell lymphoma, and bulky disease NHL;   (b) chronic lymphocytic leukemia selected from immunoglobulin-mutated CLL and immunoglobulin-unmutated CLL; and   (c) diffuse large B cell lymphoma (DLBCL) selected from germinal center B cell-like (GCB) DLBCL, activated B cell-like (ABC) DLBCL, and type 3 DLBCL.   
     
     
         82 . The method of  claim 77 , wherein administering the formulation results in the depletion of pre-B cells that do not express surface immunoglobulin by at least 50% in comparison to B cell levels in the subject before administration of the formulation. 
     
     
         83 . The method of  claim 77 , wherein administering the formulation results in the depletion of mature B cells that express surface immunoglobulin by at least 50% in comparison to B cell levels in the subject before administration of the formulation. 
     
     
         84 . The method of  claim 77 , wherein administering the formulation results in the depletion of all non-malignant B cell types by at least 50% in comparison to B cell levels in the subject before administration of the formulation. 
     
     
         85 . The method of  claim 77 , comprising administering the formulation to the subject for a period of at least about 1, 2, 3, 5, 7, 10, 14, 20, 30, 45, 60, 75, 90, 120, 150 or 180 days or longer. 
     
     
         86 . The method of  claim 77 , wherein the formulation is administered as a continuous infusion. 
     
     
         87 . The method of  claim 77 , wherein the first binding domain comprises an amino acid sequence of residues 283-525 of SEQ ID NO:5. 
     
     
         88 . The method of  claim 77 , wherein the second binding domain comprises an amino acid sequence of residues 28-277 of SEQ ID NO:5. 
     
     
         89 . The method of  claim 77 , wherein the bispecific antibody comprises an amino acid sequence of residues 28-525 of SEQ ID NO:5. 
     
     
         90 . The method of  claim 77 , wherein the formulation comprises between about 1 microgram/ml and about 500 micrograms/ml of the bispecific antibody. 
     
     
         91 . The method of  claim 77 , wherein the formulation comprises lysine. 
     
     
         92 . The method of  claim 91 , wherein the formulation further comprises:
 (a) a buffering agent selected from the group consisting of: histidine, citrate, phosphate, glycine, and acetate;   (b) a carbohydrate excipient selected from the group consisting of: sucrose, trehalose, lactose, mannitol, and raffinose; and   (c) a surfactant selected from the group consisting of: Polysorbate 20, Polysorbate 40, Polysorbate 60, and Polysorbate 80.   
     
     
         93 . The method of  claim 92 , wherein the formulation comprises citrate, trehalose, and Polysorbate 80. 
     
     
         94 . The method of  claim 77 , wherein the formulation has a pH of between about 4.0 and about 8.0 and comprises:
 (a) between about 1 micrograms/ml and about 500 micrograms/ml of the bispecific antibody;   (c) between about 5 mM and about 125 mM citrate,   (d) between about 25 mM and about 400 mM lysine,   (e) between about 3% and about 50% trehalose, and   (f) between about 0.01% and about 1% Polysorbate 80.