Novel human control antibodies and uses therefor

Abstract

The present invention provides novel, rationally designed human control antibodies for use in various in vivo and in vitro applications. The antibodies of the present invention have well characterized variable domains that have been designed to minimize or eliminate antigen binding without altering gross antibody structure. Using the antibodies of the present invention in various assays allows researchers to distinguish effects that result from specific antigen-antibody interactions from other, non-specific antibody effects.

1 . An isolated antibody or antigen binding fragment thereof, comprising: a bevacizumab reference antibody having a modification within heavy chain complementarity determining region (CDRH) 1 that eliminates target antigen binding to VEGF, wherein the CDRH1 is SEQ ID NO: 2. 
     
     
         2 . The isolated antibody or antigen binding fragment of  claim 1 , further comprising at least one more modification of heavy chain complementarity determining regions. 
     
     
         3 . An isolated antibody or antigen binding fragment thereof comprising: a bevacizumab reference antibody having a modification within CDRH2 that eliminates target antigen binding to VEGF, wherein the CDRH2 is SEQ ID NO: 4. 
     
     
         4 . An isolated antibody or antigen binding fragment thereof, comprising: a bevacizumab reference antibody having a modification within the CDRH2 that eliminates target antigen binding to VEGF, wherein the CDRH2 is SEQ ID NO: 5. 
     
     
         5 . An isolated antibody or antigen binding fragment thereof, comprising: a bevacizumab reference antibody having a modification within the CDRH3 that eliminates target antigen binding to VEGF, wherein the CDRH3 is SEQ ID NO: 7. 
     
     
         6 . The isolated antibody or antigen binding fragment thereof of  claim 2 , wherein the CDRH1 is SEQ ID NO: 2 and the CDRH3 is SEQ ID NO: 7. 
     
     
         7 . An isolated antibody or antigen binding fragment thereof, comprising: a bevacizumab reference antibody having a modification within the heavy chain variable region that eliminates target antigen binding to VEGF, wherein the heavy chain variable region has an amino acid sequence as set forth in any of SEQ ID NOs: 20-30. 
     
     
         8 . The isolated antibody or antigen binding fragment thereof of  claim 7 , wherein the isolated antibody or antigen binding fragment thereof exhibits at least 50 fold reduced specific binding affinity to the target antigen as compared to the bevacizumab reference antibody. 
     
     
         9 . The isolated antibody or antigen binding fragment thereof of  claim 7 , wherein the isolated antibody or antigen binding fragment thereof exhibits substantially no detectable specific binding affinity to the target antigen as compared to the bevacizumab reference antibody. 
     
     
         10 . The isolated antibody or antigen binding fragment thereof of  claim 7 , wherein the isolated antibody or antigen binding fragment thereof having a dissociation constant (K.sub.D) of greater than or equal to about 10.sup.-7 relative to the bevacizumab reference antibody. 
     
     
         11 . The isolated antibody or antigen binding fragment thereof of  claim 7 , wherein the isolated antibody or antigen binding fragment thereof is conjugated to a detectable moiety, a therapeutic agent, or a cytotoxic agent. 
     
     
         12 . The isolated antibody or antigen binding fragment thereof of  claim 7 , wherein the isolated antibody or antigen binding fragment thereof is selected from the group consisting of: (a) a whole immunoglobulin molecule; (b) an scFv; (c) an Fab fragment; (d) an Fab′ fragment; (e) an F(ab′)2; (f) an Fv; (g) an Fd fragment; and (h) a disulfide linked Fv. 
     
     
         13 . The isolated antibody or antigen binding fragment of  claim 1 , having modifications within the CDRH1 comprising: amino acid substitutions N50A and Y51A. 
     
     
         14 . The isolated antibody or antigen binding fragment of  claim 3 , having modifications within the CDRH2 comprising: amino acid substitutions N71A, Y73A and Y79A. 
     
     
         15 . The isolated antibody or antigen binding fragment of  claim 4 , having modifications within the CDRH2 comprising: amino acid substitutions N71A, Y73A. 
     
     
         16 . The isolated antibody or antigen binding fragment of  claim 5 , having modifications within the CDRH3 comprising: amino acid substitutions Y118A, Y121A, Y122A, S124A, S125A. 
     
     
         17 . The isolated antibody or antigen binding fragment of  claim 6 , having modifications in the CDRH1, comprising amino acid substitutions N50A and Y51A; and modifications in the CDRH3, comprising amino acid substitutions Y118A, Y121A, Y122A, S124A, S125A.