Properties of Novel Anti-digoxin Antisera in Radioimmunoassay Using Homologous and Site Heterologous Tritium-Labeled Antigens Involving a [^3H]-Leucine Moiety(Analytical Biochemistry)

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The specificities of antisera against digoxin C-3' or C-3" hemisuccinate-bovine serum albumin (BSA) conjugate were assessed by cross-reactivity studies with digoxin metabolites by radioimmunoassay (RIA) using the homologous and the site heterologous tritium-labeled antigens. One of the tracers used was digoxin 3'-hemisuccinyl-[^3H]-leucine ; the other was digoxin 3"-hemisuccinyl-[^3H]-leucine, which had been prepared from digoxin 3"-hemisuccinate. When the tracer with [^3H]-leucine at the C-3' position was used, antisera (I-1, I-3) elicited by digoxin 3'-hemisuccinate-BSA conjugate showed the following cross-reactivity : digoxigenin bisdigitoxoside (0.34%, 76%), digoxigenin monodigitoxoside (0.11%, 65%), digoxigenin (0.02%, 26%) and dihydrodigoxin (9.4%, 1.2%). However, when using the homologous antigen, antiserum (I-1) was highly specific against the digitoxose chain. When the site heterologous antigen, digoxin 3"-hemisuccinyl-[^3H]-leucine was combined, this antiserum showed high cross-reactivity to digoxin degradation products. This digoxin RIA using antiserum (I-1) with the homologous antigen measures unmetabolized digoxin. On the other hand, the RIA system using antiserum (I-3) with the homologous antigen had cross-reactivity with the metabolites in accordance with their relative cardio-activities, so this system would be useful in therapeutic drug monitoring of digoxin.
公益社団法人日本薬学会の論文
2005-02-01