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 method of treating canine lymphoma comprising administering a therapeutically effective amount of an antibody which binds to canine lymphocytes. 
     
     
         24 . A method of  claim 23 , wherein said antibody is a heterochimeric antibody, or fragment thereof, comprising amino acid sequences of antibodies from a donor species of mammal and from a target species of a different mammal, comprising:
 a) a constant region sequence from a heavy chain and light chain identical to, or a conservative variant of, a sequence from the target species; and   b) a variable domain sequence from a heavy chain and light chain, wherein said variable domain sequence comprises:
 i.) Hypervariable region sequences comprising three complementarity determing regions (CDRs) as definied by Kabat, which are identical to, or a conservative variant of, CDRs from the donor species, and 
 ii.) FR1, FR2, FR3, and FR4 framework sequences, wherein the FR1 and/or the FR4 sequence is identical to, or a conservative variant of, a FR1 and/or FR4 sequence from the target species, and the FR2 and FR3 sequences are identical to, or a conservative variant of, FR2 and FR3 sequences from the donor species; 
   wherein said variable domain sequence comprises at least three contiguous non-CDR residues corresponding to residues from the target species and at least three contiguous non-CDR residues corresponding to residues from the donor species.   
     
     
         25 . A method of  claim 23 , wherein said antibody binds to canine CD20 or CD52. 
     
     
         26 . A method of treating a patient suffering from a disease or condition characterized by the presence of abnormal cells expressing a target antigen comprising administering a therapeutically effective amount of an antibody binding to such target antigen wherein the antibody is a heterochimeric antibody, or fragment thereof, comprising amino acid sequences of antibodies from a donor species of mammal and from a target species of a different mammal, comprising:
 a) a constant region sequence from a heavy chain and light chain identical to, or a conservative variant of, a sequence from the target species; and   b) a variable domain sequence from a heavy chain and light chain, wherein said variable domain sequence comprises:
 i.) Hypervariable region sequences comprising three complementarity determing regions (CDRs) as definied by Kabat, which are identical to, or a conservative variant of, CDRs from the donor species, and 
 ii.) FR1, FR2, FR3, and FR4 framework sequences, wherein the FR1 and/or the FR4 sequence is identical to, or a conservative variant of, a FR1 and/or FR4 sequence from the target species, and the FR2 and FR3 sequences are identical to, or a conservative variant of, FR2 and FR3 sequences from the donor species; 
   wherein said variable domain sequence comprises at least three contiguous non-CDR residues corresponding to residues from the target species and at least three contiguous non-CDR residues corresponding to residues from the donor species.   
     
     
         27 . The method of  claim 26 , wherein said canine antibody binds to canine CD20. 
     
     
         28 . The method of  claim 26 , wherein said canine antibody binds to canine CD52. 
     
     
         29 . The method of  claim 26 , wherein the disease or condition characterized by the presence of abnormal cells overexpressing CD20. 
     
     
         30 . The method of  claim 26 , wherein the disease or condition characterized by the presence of abnormal cells expressing a target antigen is selected from the group consisting of: acute inflammation, rheumatoid arthritis, transplant rejection, asthma, allergic inflammation, restenosis, arterial restenosis, inflammatory bowel disease, uveitis multiple sclerosis, psoriasis, wound healing, lupus erythematosus, allergic rhinitis, atopic dermatitis, food allergies, diabetes mellitus, dermatitis, thrombotic, thrombocytopenic purpura, encephalitis, leukocyte adhesion deficiency, rheumatic fever, psoriatic arthritis, osteoarthritis, ocular inflammatory disorders, progressive systemic sclerosis, primary biliary cirrhosis, CNS inflammatory disorder, antigen-antibody complex mediated diseases, autoimmune hemolytic anemia, ischemic heart disease, atherosclerosis, post-dialysis syndrome, leukemia, acquired immune deficiency syndrome, septic shock, lipid histiocytosis, and cancer. 
     
     
         31 . The method of  claim 26 , further comprising the administration of chemotherapy. 
     
     
         32 . The method of  claim 31 , wherein the chemotherapy comprises administration of one or more agents selected from cyclophosphamide, doxorubicin, vincristine, prednisone, L-asparaginase, and adriamycin. 
     
     
         33 . The method of  claim 26 , further comprising the administration of an anti-inflammatory agent. 
     
     
         34 . The method of  claim 33 , wherein the anti-inflammatory agent is a corticosteroid. 
     
     
         35 . The method of  claim 26 , further comprising the administration of radiation. 
     
     
         36 . The method of  claim 26 , comprising the co-administration of antibody to CD20 and CD52. 
     
     
         37 . The method of  claim 26 , wherein the antibody is administered to treat or inhibit recurrence of cancer following treatment with radiation or chemotherapy. 
     
     
         38 . The method of  claim 26 , wherein the disease is canine lymphoma and the heterochimeric antibody binds to an epitope on the extracellular loop of canine CD52. 
     
     
         39 . The method of  claim 38 , further comprising the administration of a therapeutically effective amount of a heterochimeric antibody to canine CD20. 
     
     
         40 . The method of  claim 26 , wherein the heterochimeric antibody comprises a light and heavy chain, wherein the light chain is selected from the group of antibody variable domains consisting of:
 FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 T-Lambda -C T-Lambda ;   FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 T-Kappa -C T-Lambda ;   FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 T-Lambda -C T-Kappa ;   FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 T-kappa -C T-Kappa ;   FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 T-kappa -C T-Kappa ;   FR1 T-Kappa -CDR1-FR2-CDR2-FR3-CDR3-FR 4- C T-Lambda ;   FR1 T-Lambda -CDR1-FR2-CDR2-FR3-CDR3-FR4-C T-Kappa ;   FR1 T-kappa -CDR1-FR2-CDR2-FR3-CDR3-FR4-C T-Kappa ;   FR1 T-Lambda -CDR1-FR2-CDR2-FR3-CDR3-FR4 T-Lambda -C T-Lambda ; and   FR1 T-kappa -CDR1-FR2-CDR2-FR3-CDR3-FR4 T-kappa -C T-Kappa ;   wherein T=Target species; Lambda=lambda light chain; Kappa=kappa light chain; C=Constant domain; FR=Framework region; CDR=Complementarity Determining Region; and   wherein a FR is a donor species unless otherwise marked as a target species.   
     
     
         41 . The method of  claim 26 , wherein the heterochimeric antibody comprises a light and heavy chain, wherein the light chain is selected from the group of antibody variable domains consisting of:
 FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 T -C T ;   FR1 T -CDR1-FR2-CDR2-FR3-CDR3-FR4-C T ; and   FR1 T -CDR1-FR2-CDR2-FR3-CDR3-FR4 T -C T ;   wherein T=Target species; Lambda=lambda light chain; Kappa=kappa light chain; C=Constant domain; FR=Framework region; CDR=Complementarity Determining Region; and   wherein a FR is a donor species unless otherwise marked as a target species.   
     
     
         42 . The method of  claim 40 , wherein said heterochimeric antibody is an anti-CD20 or anti-CD52 monoclonal antibody. 
     
     
         43 . The method of  claim 41 , wherein said heterochimeric antibody is an anti-CD20 or anti-CD52 monoclonal antibody. 
     
     
         44 . The method of  claim 23 , wherein said antibody binds to canine CD20, and wherein the canine CD20 comprises SEQ ID NO: 67.