Anti-CD19 Antibodies

Abstract

The present invention provides humanized, chimeric and human anti-CD19 antibodies, anti-CD19 antibody fusion proteins, and fragments thereof that bind to a human B cell marker. Such antibodies, fusion proteins and fragments thereof are useful for the treatment and diagnosis of various B-cell disorders, including B-cell malignancies and autoimmune diseases. In more particular embodiments, the humanized anti-CD19 antibodies may comprise one or more framework region amino acid substitutions designed to improve protein stability, antibody binding and/or expression levels. In a particularly preferred embodiment, the substitutions comprise a Ser9lPhe substitution in the hA19 VH sequence.

What is claimed is: 
     
         1 . A method of delivering a diagnostic or therapeutic agent to a cell that expresses CD19 comprising:
 a) obtaining a humanized antibody or antigen-binding fragment thereof that binds CD19, wherein the humanized antibody or antigen-binding fragment thereof comprises (i) the light chain complementarity determining region CDR sequences CDR1 KASQSVDYDGDSYLN (SEQ ID NO: 16); CDR2 DASNLVS (SEQ ID NO: 17); and CDR3 QQSTEDPWT (SEQ ID NO: 18); (ii) the heavy chain CDR sequences CDR1 SYWMN (SEQ ID NO: 19); CDR2 QIWPGDGDTNYNGKFKG (SEQ ID NO: 20) and CDR3 RETTTVGRYYYAMDY (SEQ ID NO: 21); (iii) one or more framework region amino acid residues substituted from the corresponding framework region sequences of the parent murine antibody, wherein said one or more substituted FR residues comprise the substitution of phenylalanine for serine at Kabat residue 91 of the heavy chain variable region; and (iv) a diagnostic or therapeutic agent conjugated to said humanized antibody or antigen-binding fragment thereof to form an immunoconjugate; and   b) administering the immunoconjugate to a subject.   
     
     
         2 . The method of  claim 1 , wherein the antibody or antigen-binding fragment comprises the sequences of hA19VK (SEQ ID NO:7) and hA19VH (SEQ ID NO:10). 
     
     
         3 . The method of  claim 1 , wherein the substitution of phenylalanine for serine at VH Kabat residue 91 results in an increase in expression level of the anti-CD19 antibody in cell culture. 
     
     
         4 . The method of  claim 1 , wherein the substitution of phenylalanine for serine at VH Kabat residue 91 results in a 10-fold increase in expression level of the anti-CD19 antibody in cell culture. 
     
     
         5 . The method of  claim 1 , wherein said therapeutic agent is selected from the group consisting of a cytotoxic agent, a radionuclide, an immunomodulator, a hormone, an enzyme, an oligonucleotide and a photoactive therapeutic agent. 
     
     
         6 . The method of  claim 5 , wherein said cytotoxic agent is a drug or toxin. 
     
     
         7 . The method of  claim 6 , wherein said drug is selected from the group consisting of a vinca alkaloid, an anthracycline, a camptothecan, an epipodophyllotoxin, a taxane, a proteosome inhibitor, a nitrogen mustard, an alkyl sulfonate, a nitrosourea, an antimetabolite, an alkylating agent, a triazene, a folic acid analog, a COX-2 inhibitor, a pyrimidine analog, a purine analog, a platinum coordination complex, an antibiotic, a COX-2 inhibitor, an anti-mitotic agent, an anti-angiogenic agent and a pro-apoptotic agent. 
     
     
         8 . The method of  claim 7 , wherein said drug is selected from the group consisting of doxorubicin, methotrexate, paclitaxel, cyclophosphamide, etoposide, carmustine, vincristine, procarbazine, prednisone, bleomycin, leucovorin, phenyl butyrate, bryostatin-1 and CPT-11. 
     
     
         9 . The method of  claim 6 , wherein said toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, onconase, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin and Pseudomonas endotoxin. 
     
     
         10 . The method of  claim 5 , wherein said immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, a hematopoietic factor, a colony stimulating factor (CSF), an interferon (IFN), a stem cell growth factor, erythropoietin and thrombopoietin. 
     
     
         11 . The method of  claim 10 , wherein said lymphotoxin is tumor necrosis factor (TNF), said hematopoietic factor is an interleukin (IL), said colony stimulating factor is granulocyte-colony stimulating factor (G-CSF) or granulocyte macrophage-colony stimulating factor (GM-CSF), said interferon is interferon-alpha, -beta or -gamma, and said stem cell growth factor is 51 factor. 
     
     
         12 . The method of  claim 5 , wherein said radionuclide is selected from the group consisting of  225 Ac,  67 Ga,  90 Y,  131 I,  125 I,  186 Re,  188 Re,  177 Lu,  32 P,  64 Cu,  67 Cu,  212 Bi ,  213  Bi and  211  At 
     
     
         13 . The method of  claim 1 , wherein the humanized anti-CD19 MAb or fragment thereof is part of a bispecific or multispecific antibody or antigen-binding fragment thereof that contains a second antibody or antigen-binding fragment thereof. 
     
     
         14 . The method of  claim 9 , wherein the second antibody or fragment thereof binds to a tumor-associated antigen. 
     
     
         15 . The method of  claim 9 , wherein the second antibody or fragment thereof binds to an antigen selected from the group consisting of CD3, CD4, CDS, CD8, CD14, CD15, CD19, CD20, CD21, CD22, CD23, CD25, CD33, CD37, CD38, CD40, CD40L, CD46, CD52, CD54, CD66(a-d), CD74, CD80, CD126, CD138, B7, MUC, Ia, HLA-DR, tenascin, VEGF, P1GF, ED-B fibronectin, an oncogene product, IL-2, IL-6, TRAIL-R1 and TRAIL-R2.