Radioimmunoassay for metabolites of 9,3"-diacetylmidecamycin, a macrolide antibiotic.
A radioimmunoassay system has been developed for the measurement of two major metabolites of 9,3"-diacetylmidecamycin, Mb-6 and Mb-12. A radioimmunoassay for Mb-6 was performed by using anti-Mb-6 serum and a [125I]tyramined Mb-6 derivative as a radiolabeled antigen. The labeled antigen was prepared by the chloramine T method. The antiserum was obtained from a rabbit immunized with Mb-6 conjugated to bovine serum albumin. The obtained antiserum was cross-reactive with two other metabolites of 9,3"-diacetylmidecamycin, Mb-2 and Mb-12, in addition to Mb-6. This Mb-6 radioimmunoassay system could detect Mb-6 concentrations as low as 100 pg/ml of serum. The coefficients of variation were 4.5% (intra-assay) and 5.1% (inter-assay). A radioimmunoassay for Mb-12, using anti-midecamycin serum and a [125I]tyramined-Mb-12 derivative, has also been developed. The antiserum was cross-reactive only with Mb-12 among the 9,3"-diacetylmidecamycin metabolites. This Mb-12 radioimmunoassay system could detect Mb-12 concentrations as low as 2 ng/ml. The intra- and inter-assay variances were 5.9 and 5.8%, respectively.
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D000918:Antibody Specificity; D003429:Cross Reactions; D006801:Humans; D007457:Iodine Radioisotopes; D007933:Leucomycins / Q000378:metabolism*; D015644:Miocamycin; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods; D012979:Sodium Radioisotopes
D000900:Anti-Bacterial Agents; D007457:Iodine Radioisotopes; D007933:Leucomycins; D012979:Sodium Radioisotopes; D015644:Miocamycin