Measurement of PGE2 as the methyl oxime by radioimmunoassay using a novel iodinated label

A radioimmunoassay has been developed for prostaglandin E2 (PGE2) using methyl oxime (MOX) derivatisation and a novel 125Iodine radiolabel. PGE2-methyl oxime (PGE2-MOX) is coupled through an imide linkage to proline in a pro-gly-tyr or similar peptide rather than through the conventional amide linkage to histamine or tyrosine methyl ester. The main advantage of this method is that the imide linkage in the label does not resemble the amide link used in the original antigen and the conjugate is therefore readily displaced by the natural PGE2. This overcomes the traditional difficulty encountered in hapten RIAs where the antiserum has a higher affinity for the label than it has for the compound to be measured. The assay that has been developed using these modifications and a solid-phase second antibody separation step, is both sensitive (with a lower detection limit of 0.5 pg/tube), reliable and simple and has the advantage that methyl oximation of the sample protects the PGE from degrading prior to and during the assay.