Methods and compositions for making antibodies and antibody derivatives with reduced core fucosylation

Abstract

The invention provides methods and compositions for preparing antibodies and antibody derivatives with reduced core fucosylation.

1 - 95  (canceled) 
     
     
         96 . A method for inhibiting the growth of cells expressing CD70 antigen, comprising administering to the cells a population of humanized anti-CD70 antibodies in an amount sufficient to inhibit cell growth, wherein the antibodies each comprise:
 (i) a humanized heavy chain comprising the three CDRs from SEQ ID NO:1 and a variable region framework sequence of human germline V H 1-2 or V H 1-18 and exon J H -6, provided that any of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:1,   (ii) a humanized light chain comprising the three CDRs from SEQ ID NO:2 and a variable region framework sequence of human germline V K  exon B3 and J K  exon J k −1, provided that any of positions L25 and L33 can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:2, and   (iii) an Fc domain, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation.   
     
     
         97 . The method of  claim 96 , wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation. 
     
     
         98 . The method of  claim 96 , wherein position H46 of the antibodies is occupied by the amino acid occupying the corresponding position from SEQ ID NO:1. 
     
     
         99 . The method of  claim 96 , wherein at least one of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) in the humanized heavy chain variable region of the antibodies is occupied by the residue occupying the corresponding position in SEQ ID NO:1. 
     
     
         100 . The method of  claim 96 , wherein at least one of positions L25 and L33 in the humanized light chain variable region of the antibodies is occupied by the residue occupying the corresponding position in SEQ ID NO:2. 
     
     
         101 . The method of  claim 96 , wherein the antibodies comprise a human IgG1 constant domain, wherein the humanized heavy chain comprises the amino acid sequence corresponding to positions 20-467 of SEQ ID NO:3 or positions 20-467 of SEQ ID NO:4. 
     
     
         102 . The method of  claim 96 , wherein the antibodies each comprise an antibody effector domain. 
     
     
         103 . The method of  claim 102 , wherein the antibody effector domain mediates ADCP. 
     
     
         104 . The method of  claim 96 , wherein the antibodies are conjugated to therapeutic agents. 
     
     
         105 . The method of  claim 104 , wherein the therapeutic agents are chemotherapeutic agents or immunomodulatory agents. 
     
     
         106 . A method for inhibiting the growth of cells expressing CD70 antigen, comprising administering to the cells a population of humanized anti-CD70 antibodies in an amount sufficient to inhibit cell growth, wherein the antibodies each comprise:
 (i) a humanized heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, or amino acids 20-137 of SEQ ID NO:9,   (ii) a humanized light chain variable region comprising the amino acid sequence of SEQ ID NO:10, and   (iii) an Fc domain, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation.   
     
     
         107 . The method of  claim 106 , wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation. 
     
     
         108 . The method of  claim 106 , wherein the humanized heavy chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:7 or SEQ ID NO:8, and the humanized light chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:10. 
     
     
         109 . The method of  claim 106 , wherein the humanized heavy chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:7 and the humanized light chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:10. 
     
     
         110 . The method of  claim 109 , wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation. 
     
     
         111 . The method of  claim 106 , wherein the antibodies are conjugated to therapeutic agents. 
     
     
         112 . The method of  claim 111 , wherein the therapeutic agents are chemotherapeutic agents or immunomodulatory agents. 
     
     
         113 . A method for inhibiting the growth of cells expressing CD70 antigen, comprising administering to the cells a population of humanized anti-CD70 antibodies in an amount sufficient to inhibit cell growth, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation, and wherein the antibodies are h1F6. 
     
     
         114 . The method of  claim 113 , wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation. 
     
     
         115 . The method of  claim 113 , wherein the antibodies are conjugated to therapeutic agents.