Glycosylated humanized B-cell specific antibodies

Abstract

A humanized specific monoclonal antibody or antibody fragment, especially a B-cell specific antibody or antibody fragment, is engineered to contain a glyxosylation site in the non-Fc constant region. The glycosylated antibody is useful for diagnosis and/or therapy whenever a targeting antibody or fragment is used, especially for B-cell malignancies. The carbohydrate moiety allows conjugation of labeling or therapeutic agents of increased size, without affecting the binding affinity or specificity of the antibody.

What is claimed is:  
     
         1 . A monoclonal antibody or antibody fragment which is engineered to contain a glycosylation site in the non-Fc constant heavy-chain or light-chain region.  
     
     
         2 . The humanized monoclonal antibody or antibody fragment of  claim 1 , which is a humanized antibody or antibody fragment.  
     
     
         3 . The humanized specific monoclonal antibody or fragment of  claim 2 , which is a humanized B-cell specific antibody or antibody fragment.  
     
     
         4 . The humanized B cell specific monoclonal antibody or fragment according to  claim 3 , wherein said glycosylation is located on a site selected from the group consisting of the HCN1, HCN2, HCN3, HCN4, and HCN5 sites of FIG. 12.  
     
     
         5 . The humanized B cell specific monoclonal antibody or antibody fragment according to  claim 2 , wherein said glycosylation site is the HCN5 site of FIG. 12.  
     
     
         6 . The humanized B cell specific monoclonal antibody or fragment according to  claim 2 , wherein said glycosylation site is the HCN1 site of FIG. 12.  
     
     
         7 . The antibody of  claim 3 , where the antibody which is engineered to contain a glycosylation site is an antibody having the specificity of the hLL2 antibody.  
     
     
         8 . An isolated DNA molecule comprising an antibody heavy chain gene which comprises a sequence within the CHI region that, when said gene is coexpressed with a second gene for an antibody light chain in a cell supporting glycosylation, will produce an antibody glycosylated in the CH1 region.  
     
     
         9 . An isolated DNA molecule comprising an antibody light chain gene which comprises a sequence within the constant region, which, when said gene is coexpressed with a second gene for an antibody heavy chain in a cell supporting glycosylation, will produce an antibody glycosylated in the constant K region.  
     
     
         10 . A method of producing an antibody or antibody fragment glycosylated in the constant K and/or CH1 region, comprising coexpressing light and heavy chain genes or portions thereof, which have been engineered with a mutation such that a glycosylation site is created in the constant K region or into the CH1 region of said heavy chain gene or portions thereof, in a cell that allows glycosylation, such that said antibody or antibody fragment glycosylated in the constant K and/or CH1 region is produced, and isolating said antibody or antibody fragment.  
     
     
         11 . In a method of diagnosis or treatment of a patient wherein a monoclonal antibody or antibody fragment is used to target a specific antigen, the antibody or fragment being used as such or conjugated to a diagnostic or therapeutic agent, the improvement wherein said antibody or fragment is the humanized monoclonal antibody or antibody fragment of  claim 2 .  
     
     
         12 . The method of  claim 11 , wherein said patient suffers from a B-cell malignancy and said humanized monoclonal antibody or antibody fragment is a B-cell specific antibody or antibody fragment.  
     
     
         13 . The method of  claim 11 , where said diagnostic or therapeutic agent is conjugated to a carbohydrate or said monoclonal antibody or antibody fragment.