[An improved method for determination of C-peptide levels in serum: pretreatment of test samples with anti-insulin antibody insolubilized with magnetic particles].

Abstract RIA using anti-C-peptide antibody has been employed for determination of C-peptide in serum. However, the level by this method is a sum of the levels of C-peptide and proinsulin, because anti-C-peptide cross-reacts with proinsulin. This time we developed an improved assay for C-peptide, in which a test sample is pretreated with anti-insulin antibody to eliminate proinsulin in samples before C-peptide assay. The assay procedure is composed of 3 steps; the first is the incubation of test serum with anti-insulin antibody insolubilized with magnetic particles to form a complex of proinsulin-anti-insulin antibody, the 2nd step is the centrifugation of the mixture to eliminate the complex and the last step is the assay for C-peptide by use of RIA kit in the supernatant. The assay is simple, sensitive and reproducible. Serum C-peptide level by this method is not influenced by the presence of proinsulin in test serum, even when as high as 9 ng/ml of proinsulin is contained. This assay revealed that a patient with insulinoma had normal level of serum C-peptide in spite of the extremely high level of proinsulin.