Radioimmunoassay for medroxyprogesterone acetate (Provera) using the 11alpha-hydroxy succinyl conjugate |
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Authors: | M E Royer H Ko J A Campbell H C Murray J S Evans D G Kaiser |
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Institution: | 1. Division of Reproductive Biology The Milton S. Hershey Medical Center, Hershey, Pa., USA;2. the Eleanor Roosevelt Cancer Foundation Research Institute and the Department of Biochemistry, University of Utah College of Medicine, Salt Lake City, Utah, USA |
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Abstract: | The development of a radioimmunoassay for medroxyprogesterone acetate (MPA) employing rabbit antibodies made against a bovine serum albumin (BSA) conjugate of the hemisuccinate ester of 11alpha-hydroxy MPA are described. The 11alpa-hydroxyl group was introduced into MPA by a series of chemical and microbiological transformations. The acid succinate MPA was coupled to BSA by reacting it with N,N'-carbonyldiimidazole. Cross-reactivities of several MPA analogs were compared suing rabbit antisera developed against the C-11 conjugate of MPA and goat antiserum developed against a C-3 conjugate of MPA. Antibodies formed against the C-11 conjugate showed increased specificity to steroid analogs which changes in the C-21 side chain. The profiles of serum MPA levels vs time, measured with both the C-3 and C-11 antisera, after oral drug administration to the monkey were similar. After intramuscular drug administration to dogs, serum MPA levels measured with the C-3 antisera were consistently greater as compared with those with C-11 antisera. Factors affecting precipitation of the antibody-antigen complex, assay precision, and assay sensitivity were evaluated. |
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