Enzyme immunoassay for pepleomycin, a new bleomycin analog.

An antibody directed toward pepleomycin, a new antitumor antibiotic related structurally to bleomycin, has been produced in rabbits by immunization with a pepleomycin-protein conjugate which was prepared by a novel procedure of coupling pepleomycin to mercaptosuccinylated bovine serum albumin using N-(gamma-maleimidobutyryloxy)succinimide as a coupling agent. The antiserum was monospecific to pepleomycin and showed almost no cross-reactivity with a variety of other bleomycin analogs. An enzyme immunoassay for pepleomycin has been developed utilizing this antiserum and beta-D-galactosidase-labeled pepleomycin. The lower limit of detection by this assay, which involves a double antibody technique for the separation of antibody-bound and free antigen, was 50 pg of pepleomycin per tube. Using this assay, drug levels were easily determined in blood and urine of rabbits following administration of pepleomycin in a single dose of 1.2 mg/kg i.v. This assay is also suitable for measuring pepleomycin in the presence of other drugs since the assay is not significantly affected by any other antineoplastic agents tested. Since pepleomycin is now undergoing clinical trial, the enzyme immunoassay of the drug will be a valuable tool in clinical pharmacological studies.