Development of a sensitive monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for mitozantrone.
A mouse monoclonal antibody (NO-1) with specificity for the anti-cancer drug mitozantrone (MZ) (Novantrone) was produced by immunization of a BALB/c mouse with mitozantrone-keyhole limpet haemocyanin (MZ-KLH) conjugate. When used in an indirect competitive enzyme-linked immunosorbent assay (ELISA), NO-1 permitted the accurate and reproducible detection of between 0.25-50 ng/ml of MZ in pooled human serum, the standard curve obtained within this range being virtually linear. The assay demonstrated good reproducibility with intra-assay coefficients of variation (CV) of between 1.41% and 7.02% and an inter-assay CV of 3.45%. Regression analysis of levels of MZ detected by ELISA vs. the actual amounts added to pooled human serum gave a very good correlation coefficient of r = 0.995. NO-1 showed no cross-reactivity with either bisantrene or daunorubicin. A simple pharmacokinetic study was undertaken in rabbits given MZ intravenously at a dose of 0.5 mg/kg of body weight. Levels of MZ in rabbit serum measured with the assay ranged between 82 and 170 ng/ml for rabbits 1 and 2, respectively at 15 min falling to 1.25 ng/ml by 48 h for rabbit 1 and falling to undetectable levels by 120 h for rabbit 2.
Flavell
S U
SU
University Department of Haematology, Southampton General Hospital, Hampshire, U.K.
Flavell
D J
DJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000592:standards; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002474:Cell-Free System; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D006801:Humans; D006825:Hybridomas / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D008942:Mitoxantrone / Q000097:blood* / Q000592:standards; D011817:Rabbits; D012015:Reference Standards; D012016:Reference Values; D012710:Serum Albumin, Bovine
D000911:Antibodies, Monoclonal; D012710:Serum Albumin, Bovine; D008942:Mitoxantrone