Antibodies to adenosine 5'-monophosphate: purification and specificity

Antibodies to adenosine-5'-monophosphate were produced in rabbits by injecting a conjugate of the nucleotide (oxidized with periodate) with bovine serum albumin. Nucleotide-specific antibodies were isolated by affinity chromatography on oligoadenylic acids/agarose column. Pure immunoglobulin G antibodies were obtained by gel filtration on Sephadex G-200. These antibodies, as analyzed by double diffusion react with adenosine 5'-monophosphate--bovine serum albumin, slightly with inosine-5'-monophosphate conjugate and not at all with the other nucleotide conjugates. The association constants for adenosine-5'-monophosphate--antibody complex formation obtained by dialysis equilibrium and fluorescence measurements, are in good agreement. This latter technique was used to study on one hand the influence of temperture and salt on complex formation, on the other hand the interaction of the antibodies with AMP derivatives. The phosphate group, the ribose and the base are recognized by the antibody, but the C-8 atom of adenine residues is not directly involved in the binding.