A Two-Site Immunoradiometric Assay for Serum Calcitonin Using Monoclonal Anti-Peptide Antibodies

We have produced a library of monoclonal antibodies of various affinities by immunizing mice with synthetic calcitonin (CT) 1-32. These monoclonal antibodies defined two antigenic determinants on the molecule of CT. The first was located in the 11-17 region of the hormone: the second was present on the 26-32 portion of CT. The C-terminal epitope was restricted to the mature form of the hormone and immunologically silent on synthetic peptides with sequences analogous lo the biosynthetic precursors for CT. Using two high-affinity monoclonal antibodies, designated as CT07 and CT08, we developed a two-site immunoradiometric assay (m-lRMA) for serum CT. This m-lRMA provided a sensitivity of 10 pg/mL using a one-step overnight incubation at room temperature. Gel filtration analyses of serum samples from patients with medullary thyroid carcinoma (MTC) demonstrated that the CT07-CT08 m-lRMA was specific for the circulating mature form of CT