Further production and characterization of antibodies reactive with pyrazolone derivatives

A sensitive radioimmunoassay for pyrazolone derivatives has been developed. Anti-antipyrine antisera were produced in rabbits by repeated immunization with 4-succinamidoantipyrine coupled to bovine serum albumin. Less than 1 ng of antipyrine could be detected by this procedure. Various substituents on the carbon-4 position of the pyrazolone ring decreased the affinity for the antibody. The concentrations in ng of various pyrazolone derivatives required to inhibit [3H]antipyrine binding by 50% were: antipyrine, 6.8; aminopropylon, 8.5; sulpyrine, 35.5; isopropylantipyrine, 1320; and aminopyrine, 2820. The antibody showed no cross-reactivity with any other antipyretics such as pyrazolidine or aniline derivatives. The determination of antipyrine and sulpyrine concentrations in rat serum after i.p administration was also carried out.