Introduction of nucleophiles and spectroscopic probes into antibody combining sites.
A general chemical strategy has been developed whereby antibody combining sites can be selectively derivatized with natural or synthetic molecules, such as catalytic groups, drugs, metals, or reporter molecules. Cleavable affinity labels were used to selectively introduce a thiol into the combining site of the immunoglobulin A MOPC 315. This thiol acted both as a nucleophile to accelerate ester thiolysis 60,000-fold and as a handle for selectively derivatizing the antibody with additional functional groups. For example, derivatization of the antibody with a fluorophore made possible a direct spectroscopic assay of antibody-ligand complexation. This chemistry should not only extend our ability to exploit antibody specificity in chemical catalysis, diagnostics, and therapeutics, but may also prove generally applicable to the functional modification of other proteins for which detailed structural information is unavailable.
Pollack
S J
SJ
Department of Chemistry, University of California, Berkeley 94720.
Nakayama
G R
GR
Schultz
P G
PG
D000345:Affinity Labels; D000818:Animals; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D055598:Chemical Phenomena; D002621:Chemistry; D004136:Dinitrobenzenes; D007140:Immunoglobulin Fab Fragments; D051379:Mice; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds
D000345:Affinity Labels; D004136:Dinitrobenzenes; D007140:Immunoglobulin Fab Fragments; D013438:Sulfhydryl Compounds