Clinical Chemistry
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Clinical Chemistry 38: 227-232, 1992;
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Clinical Chemistry, Vol 38, 227-232, Copyright © 1992 by American Association for Clinical Chemistry

Amplified enzyme-linked immunoassay of human proinsulin in serum (detection limit: 0.1 pmol/L)

FJ Dhahir, DB Cook and CH Self
Department of Clinical Biochemistry, Medical School, University of Newcastle upon Tyne, U.K.

We describe an amplified enzyme-linked immunoassay of human proinsulin in serum that detects intact proinsulin and both the 32/33 and 65/66 split forms. The method uses the IgG fraction of a polyclonal antibody raised in a guinea pig against intact proinsulin, which we used to coat plastic microtiter plates. A sandwich was formed with proinsulin by using a monoclonal antibody against C-peptide labeled with alkaline phosphatase. We quantified the reaction by using the enzyme amplification procedure, which detected as little intact proinsulin as 0.1 pmol/L. We found no cross-reactivity with C-peptide in the assay, and decreased recovery attributable to the presence of insulin could be demonstrated only with a 30-fold excess of this hormone over proinsulin.


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 Clin. Chem.Home page
P. Houssa, B. Dinesen, M. Deberg, B. H. Frank, C. Van Schravendijk, F. Sodoyez-Goffaux, and J.-C. Sodoyez
First direct assay for intact human proinsulin
Clin. Chem., July 1, 1998; 44(7): 1514 - 1519.
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Copyright © 1992 by the American Association for Clinical Chemistry.