The Estimation of Elastin in Fetal Tissues by Radioimmunoassay of Isodesmosine

Abstract

A radioimmunoassay was developed for the determination of isodesmosine as the tetraacetyl derivative. Iso-desmosine tetraacetate conjugated with bovine albumin was injected into rabbits which developed useful titers of antibodies after five months. The radioligand for the assay was prepared by acetylating isodesmosine with [³H] acetic anhydride. The bound ligand was separated from free ligand by coprecipitation with human γ-globulin in 46% saturated ammonium sulfate solution. The sensitivity of the assay was 2 ng isodesmosine. The antiserum was specific for isodesmosine tetraacetate and only desmosine tetraacetate gave appreciable cross-reactivity (4%). The assay was found to be suitable for the accurate estimation of elastin in small samples (5 mg dry weight) of rat and ovine fetal lung tissue and for elastin degradation products in amniotic fluid (0.5 ml).