Use of a native epitope for selecting evolved binding members from a library of mutants of a protein capable of binding to said epitope

Abstract

The invention provides a method for selecting at least one member from a library of proteinaceous molecules comprising providing at least one cell and/or a functional equivalent thererof, with at least part of said library under conditions that allow binding of any such member to an epitope in and/or on said cells and/or said functional equivalent thereof, removing unbound proteinaceous molecules and selecting said at least one member, wherein said library comprises at least one mutant of a proteinaceous molecule capable of binding to said epitope.

1 . A method for selecting at least one member from a library of proteinaceous molecules comprising providing at least one cell and/or a functional equivalent thereof, with at least part of said library under conditions that allow binding of any such member to an epitope in and/or on said cells and/or said functional equivalent thereof, removing unbound proteinaceous molecules and selecting said at least one member, wherein said library comprises at least one mutant of a proteinaceous molecule capable of binding to said epitope.  
     
     
         2 . A method according to  claim 1 , wherein at least one of said proteinaceous molecules comprises a single chain antibody and/or a FAB fragment, or a functional equivalent thereof.  
     
     
         3 . A method according to  claim 1  or  claim 2 , wherein said at least one mutant of a proteinaceous molecule is associated with nucleic acid encoding said at least one mutant proteinaceous molecule.  
     
     
         4 . A method according to  claim 3 , wherein said association is achieved through a vehicle that is physically linked to said at least one mutant proteinaceous molecule.  
     
     
         5 . A method according to  claim 4 , wherein said vehicle comprises a virus-like particle such as a phage capsid or a functional equivalent thereof.  
     
     
         6 . A method according to anyone of claims  1 - 5 , wherein said epitope comprises a tumour-associated epitope.  
     
     
         7 . A proteinaceous molecule obtainable by a method according to anyone of claims  1 - 6 .  
     
     
         8 . A molecule capable of binding to an epitope, comprising at least part of a member obtained with a method according to anyone of claims  1 - 6 .  
     
     
         9 . A molecule according to  claim 7  or  claim 8 , wherein said molecule comprises an antibody or a functional part thereof.  
     
     
         10 . A molecule according to  claim 9 , wherein said antibody is human, humanised and/or human-like, or a functional equivalent thereof.  
     
     
         11 . Use of a cell and/or a functional equivalent thereof displaying an epitope, for obtaining an evolved epitope binding molecule with an enhanced property as compared to the epitope binding molecule said evolved epitope binding molecule is at least in part derived from.  
     
     
         12 . A use according to  claim 11 , wherein said epitope binding molecule comprises a part of a complementarity determining region of an antibody or a functional equivalent thereof.  
     
     
         13 . A use according to  claim 11  or  claim 12 , wherein said property comprises an enhanced epitope binding property.  
     
     
         14 . A use according to anyone of claims  11 - 13 , wherein said property comprises an enhanced tissue penetration property.  
     
     
         15 . A use according to anyone of claims  11 - 14 , wherein said property comprises an enhanced complement activation property.