Monoclonal antibodies

Abstract

The invention provides heterochimeric antibodies and/or fragments thereof comprising (i) hypervariable region sequences wholly or substantially corresponding to sequences found in antibodies from a donor species; (ii) constant region sequences wholly or substantially corresponding to sequences found in antibodies from a target species which is different from the donor species; and (iii) heavy and/or light chain variable framework sequences which contain at least three non-CDR residues corresponding to sequences found in antibodies from a target species and at least three contiguous non-CDR residues corresponding to sequences found in antibodies from a donor species. The invention further provides antibody to canine or feline or equine antigens, e.g., CD20 or CD52, and methods of making and using antibodies as described.

1 .- 22 . (canceled) 
     
     
         23 . A canine CD20 antigen comprising an extracellular domain and wherein the extracellular domain comprises an epitope, wherein the epitope comprises a sequence selected from the group consisting of SEQ ID NOS 78, 81, 82, and 83. 
     
     
         24 . The canine CD20 antigen of  claim 23 , wherein the extracellular domain comprises SEQ ID NO: 75. 
     
     
         25 . The CD20 antigen of  claim 23 , wherein the canine CD20 antigen comprises SEQ ID NO: 67. 
     
     
         26 . An antibody which binds to an epitope on a extracellular loop of canine CD20, wherein said epitope comprises a sequence selected from SEQ ID NOS 78, 81, 82, and 83. 
     
     
         27 . The antibody according to  claim 26 , wherein the extracellular domain comprises SEQ ID No. 75. 
     
     
         28 . The antibody according to  claim 26  which is derived from or has substantially the same hypervariable domain as an antibody raised against an immunogenic construct comprising or expressing a peptide comprising a sequence selected from one or more of SEQ ID NOS. 75, 78, 81, 82, and 83. 
     
     
         29 . The antibody according to  claim 26  wherein the antibody comprises hypervariable sequences from a donor species antibody and constant region sequences from a target species, wherein target species is canine. 
     
     
         30 . The antibody according to  claim 29  wherein the antibody is caninized. 
     
     
         31 . The antibody according to  claim 26 , wherein the canine CD20 comprises SEQ ID NO: 67. 
     
     
         32 . The antibody of  claim 26 , wherein the antibody causes the death of cells expressing CD20 by antibody dependent cell-mediated cytotoxicity (ADCC) 
     
     
         33 . The antibody of  claim 26 , wherein the antibody causes the death of cells expressing CD20 by antibody complement dependent cytotoxicity (CDC) 
     
     
         34 . An immunogenic construct comprising or expressing a peptide comprising a sequence selected from one or more of SEQ ID NOS. 75, 78, 81, 82, and 83. 
     
     
         35 . An immunogenic construct according to  claim 34 , wherein the peptide comprises SEQ ID NO: 67. 
     
     
         36 . A method of making an antibody according  claim 26 , wherein the method comprises raising an antibody against an immunogenic construct comprising or expressing a peptide comprising a sequence selected from one or more of SEQ ID NOS. 75, 78, 81, 82, and 83. 
     
     
         37 . The method according to  claim 36 , wherein the peptide comprises SEQ ID NO: 67. 
     
     
         38 . A method of a method of treating a dog suffering from a disease or condition characterized by the presence of abnormal cells expressing CD20 comprising administering a therapeutically effective amount of an antibody according to  claim 26 . 
     
     
         39 . The method of  claim 38 , wherein the condition to be treated is canine lymphoma. 
     
     
         40 . The method of  claim 39 , further comprising administration of an additional therapy, wherein the therapy is selected from the group consisting of: chemotherapy, a corticosteroid, and radiation. 
     
     
         41 . The method of  claim 40 , wherein the additional therapy is radiation. 
     
     
         42 . The method of  claim 40 , wherein the additional therapy is the administration of a corticosteroid. 
     
     
         43 . The method of  claim 40 , wherein the additional therapy is chemotherapy. 
     
     
         44 . The method of  claim 43 , wherein the chemotherapy comprises administration of one or more agents selected from the group consisting of: cyclophosphamide, doxorubicin, vincristine, prednisone, L-asparaginase, cytoxan and adriamycin. 
     
     
         45 . The method of  claim 38 , further comprising the co-administration of an antibody to CD52. 
     
     
         46 . A method to treat or inhibit recurrence of cancer comprising administration of the antibody of  claim 26  following treatment with radiation or chemotherapy. 
     
     
         47 . The method of  claim 46 , wherein the antibody binds to a canine CD20 which comprises SEQ ID NO: 67. 
     
     
         48 . A pharmaceutical composition comprising the antibody of  claim 26 . 
     
     
         49 . A cell line stably expressing an antibody according to  claim 26 . 
     
     
         50 . The cell line of  claim 49  which is a CHO cell line. 
     
     
         51 . A vector or vectors expressing at least one heavy chain and at least one light chain of an antibody according to any of  claims 26 . 
     
     
         52 . A method of making an antibody comprising transforming a cell line with a vector or vectors expressing at least one heavy chain and at least one light chain of an antibody according to any of  claim 26 . 
     
     
         53 . The method according to  claim 52 , wherein the vector or vectors encode the full-length CD20 protein, wherein said protein comprises SEQ ID NO: 67.