Regioselective labeling of antibodies through N-terminal transamination.
A convenient new method is described for the introduction of ketone groups at the N-termini of antibodies. The reaction occurs in the presence of pyridoxal-5'-phosphate under conditions mild enough to maintain antigen binding function, as confirmed by enzyme-linked immunosorbent assay. Further derivatization of these functional sites was accomplished through oxime formation, yielding well-defined antibody conjugates for a wide range of applications. The ability of the modified antibodies to bind their targets was confirmed via immunodot blot analysis. The generality of this method has been demonstrated on a number of monoclonal and polyclonal antibodies, all with different binding specificities.
Scheck
Rebecca A
RA
Francis
Matthew B
MB
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry; D007659:Ketones / Q000737:chemistry; D051379:Mice; D010091:Oximes / Q000737:chemistry; D011732:Pyridoxal Phosphate / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007659:Ketones; D010091:Oximes; D011732:Pyridoxal Phosphate