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Clinical Chemistry 44: 1504-1513, 1998;
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(Clinical Chemistry. 1998;44:1504-1513.)
© 1998 American Association for Clinical Chemistry, Inc.

Endocrinology and Metabolism

Highly specific radioimmunoassay for human insulin based on immune exclusion of all insulin precursors

Michelle Deberg1, Paule Houssa1, Bruce H. Frank2, Françoise Sodoyez-Goffaux1,a, and Jean-Claude Sodoyez1,2

1 University of Liège, Division of Nuclear Pediatrics, Sart Tilman, 4000 Liège, Belgium.

2 Lilly Research Laboratories, 307 East Mccarty Street, Indianapolis, IN 46285.
a Author for correspondence. Fax 32-4-366-82-55.

We describe a rapid and simple insulin RIA in which proinsulin and conversion intermediates do not interfere. Three monoclonal antibodies (S1, S2, and S53) were selected for their specificity (directed, respectively, against the B10 region, the junction between A chain and C-peptide, and the junction between B chain and C-peptide), their affinity constant (~1010 L/mol), and their interactive properties in mixture. S2 and S53 were able to bind simultaneously to the same proinsulin molecule, whereas neither could bind simultaneously with S1. Preincubation of serum samples with an excess of S2 resulted in capture of proinsulin and conversion intermediates modified at the junction between B chain and C-peptide into immune complexes that no longer reacted with S1. Similarly, preincubation with S53 prevented proinsulin and conversion intermediates modified at the junction between A chain and C-peptide from reacting with S1. Preincubation with an excess of both S2 and S53 left insulin as the sole reactant with S1. Thus, separation of insulin precursors from insulin by mutually exclusive antibodies is feasible, and on the basis of this new principle, a highly specific RIA for insulin was designed. The detection limit was 11 pmol/L, and the inter- and intraassay coefficients of variation were 11% and 5%, respectively. The potential of the assay for use in clinical studies was verified by application to serum samples from control subjects and patients with diabetes or insulinoma.




The following articles in journals at HighWire Press have cited this article:


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 Clin. Chem.Home page
D. Rodriguez-Cabaleiro, K. Van Uytfanghe, V. Stove, T. Fiers, and L. M. Thienpont
Pilot Study for the Standardization of Insulin Immunoassays with Isotope Dilution Liquid Chromatography/Tandem Mass Spectrometry
Clin. Chem., August 1, 2007; 53(8): 1462 - 1469.
[Abstract] [Full Text] [PDF]

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C. W. Chia and C. D. Saudek
The Diagnosis of Fasting Hypoglycemia Due to an Islet-Cell Tumor Obscured by a Highly Specific Insulin Assay
J. Clin. Endocrinol. Metab., April 1, 2003; 88(4): 1464 - 1467.
[Abstract] [Full Text] [PDF]


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