Engineering antibodies that bind irreversibly

A polypeptide comprising a peptide sequence according to SEQ. ID NO.:5 (FIG. 12).

A polypeptide comprising a peptide sequence according to SEQ. ID NO.:

• 7 (FIG. 14).

A mutant antibody comprising a reactive site containing a cysteine residue not present in the wild-type of said antibody and six complementarity determining regions (CDRs) that recognize a metal chelate or portions thereof, wherein said cysteine is in a position proximate to or within said complementarity-determining regions, wherein said cysteine residue is the mutation and wherein said reactive site interacts with a reactive group on said metal chelate.

A mutant antibody comprising a reactive site not present in the wild-type of said antibody and six complementarity determining regions (CDRs) that specifically bind a metal chelate, wherein said reactive site is in a position proximate to or within said complementarity-determining regions,

• wherein said reactive site is the mutation and,
• wherein said reactive site interacts with a reactive group on the metal chelate selected from carboxyl groups, hydroxyl groups, haloalkyl groups, dienophile groups, aldehyde groups, ketone groups, sulfonyl halide groups, thiol groups, amine groups, sulfhydryl groups, alkene groups, and epoxide groups.

A mutant antibody comprising a reactive site not present in the wild-type of said antibody and six complementarity determining regions (CDRs) that recognize a metal chelate or portions thereof, wherein said reactive site is in a position proximate to or within said complementarity-determining regions,

• wherein said reactive site is introduced by mutagenizing a nucleic acid encoding said wild-type of said antibody and,
• wherein said reactive site interacts with a reactive group on said metal chelate and said reactive group is selected from carboxyl groups, hydroxyl groups, haloalkyl groups, dienophile groups, aldehyde groups, ketone groups, sulfonyl halide groups, thiol groups, amine groups, sulfhydryl groups, alkene groups, and epoxide groups.

A mutant antibody comprising a reactive site not present in the wild-type of said antibody and six complementarity determining regions (CDRs) that specifically bind a metal chelate, wherein said reactive site is in a position proximate to or within said complementarity-determining regions,

• wherein said reactive site is introduced by mutagenizing a nucleic acid encoding said wild-type of said antibody, and
• wherein said reactive site interacts with a reactive group on the metal chelate selected from carboxyl groups, hydroxyl groups, haloalkyl groups, dienophile groups, aldehyde groups, ketone groups, sulfonyl halide groups, thiol groups, amine groups, sulfhydryl groups, alkene groups, and epoxide groups.