Production of humanized antibodies in transgenic animals

Abstract

This invention relates to humanized antibodies and antibody preparations produced from transgenic non-human animals. The non-human animals are genetically engineered to contain one or more humanized immunoglobulin loci which are capable of undergoing gene rearrangement and gene conversion in the transgenic non-human animals to produce diversified humanized immunoglobulins. The present invention further relates to novel sequences, recombination vectors and transgenic vectors useful for making these transgenic animals. The humanized antibodies of the present invention have minimal immunogenicity to humans and are appropriate for use in the therapeutic treatment of human subjects.

1 . A humanized immunoglobulin molecule comprising at least a portion of a human immunoglobulin heavy chain constant (C) region, and variable region amino acids sequences encoded by more than one immunoglobulin variable (V) gene segment.  
     
     
         2 . The humanized immunoglobulin molecule of  claim 1  which is derived from a nonhuman transgenic animal relying primarily on gene conversion and/or somatic hypermutation to generate antibody diversity.  
     
     
         3 . The humanized immunoglobulin molecule of  claim 2  wherein said transgenic animal is a rabbit, chicken, sheep or cow.  
     
     
         4 . The humanized immunoglobulin molecule of  claim 1  wherein said human immunoglobulin heavy chain C region sequence is a C γ , C κ , or C λ  sequence.  
     
     
         5 . The humanized immunoglobulin molecule of any one of claims  1 - 4 , which is specific for an antigen.  
     
     
         6 . The humanized immunoglobulin molecule of  claim 5 , wherein said antigen is a microorganism selected from bacterium, fungus, or virus; an antigenic portion of said organism; an antigenic molecule derived from said microorganism; or a tumor-associated antigen.  
     
     
         7 . The humanized immunoglobulin molecule of  claim 6 , wherein said bacterium is selected from  S. aureus, Pseudomonas aeruginosa, enterococcus, enterobacter , and  Klebsiella pneumoniae.    
     
     
         8 . The humanized immunoglobulin molecule of  claim 6 , wherein said fungus is selected from  Candida albicans, Candida parapsilosis, Candida tropicalis , and  Cryptococcus neoformans.    
     
     
         9 . The humanized immunoglobulin molecule of  claim 6 , wherein said virus is selected from respiratory synctial virus (RSV), Hepatitis C virus (HCV), Hepatitis virus (HBV), cytomegalovirus (CMV), EBV, and HSV.  
     
     
         10 . The humanized immunoglobulin molecule of  claim 6 , wherein said antigen is selected from Her-2-neu antigen, CD20, CD22, CD53, prostate specific membrane antigen (PMSA), and 17-1A molecule.  
     
     
         11 . A monoclonal antibody preparation comprising a humanized immunoglobulin molecule of  claim 5 .  
     
     
         12 . A monoclonal antibody preparation comprising a humanized immunoglobulin molecule of any one of claims  6 - 10 .  
     
     
         13 . The monoclonal antibody preparation of  claim 11  which is substantially non-immunogenic to human.  
     
     
         14 . The monoclonal antibody preparation of  claim 12  which is substantially non-immunogenic to human.  
     
     
         15 . A pharmaceutical composition, comprising a pharmaceutically acceptable carrier and the monoclonal antibody preparation of  claim 11 .  
     
     
         16 . A pharmaceutical composition, comprising a pharmaceutically acceptable carrier and the monoclonal antibody preparation of  claim 12 .  
     
     
         17 . A method of treating a disease in a human subject comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition of  claim 15 .  
     
     
         18 . The method of  claim 17  wherein said disease is caused by bacterial, fungal or viral infection, or said disease is a cancer.