Immunoassay of leucovorin: use of 125I-labeled protein A to detect immunological binding. |
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Authors: | J J Langone L Levine |
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Institution: | 1. Laboratory of Immunobiology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA;2. Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02154 USA |
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Abstract: | An immunoassay method was developed for the quantitative determination of leucovorin. Leucovorin immobilized by covalent linkage to a solid support bound anti-leucovorin antibody produced in rabbits. The amount of 125I-labeled Protein A bound specifically to IgG anti-leucovorin-coated beads served as a measure of antibody binding. The ability of increasing amounts of fluid-phase leucovorin to compete with solid-phase drug for specific antibody was reflected in a dose-dependent decrease in the amount of bound 125I-labeled Protein A. This competition was used to obtain a standard curve to measure levels of leucovorin in the sera of rabbits treated with the drug. The relative abilities of methotrexate, 5-methyltetrahydrofolic acid, and other structurally related compounds to act as inhibitors demonstrated the serologic specificity of the leucovorin immune system. |
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