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Related Experiment Videos

First direct assay for intact human proinsulin

P Houssa1, B Dinesen, M Deberg

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

Clinical Chemistry
|July 17, 1998
PubMed
Summary

A new sensitive assay accurately measures intact human proinsulin in serum or plasma. This method shows significantly elevated proinsulin levels in diabetes and insulinoma patients compared to healthy individuals.

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Area of Science:

  • Biochemistry
  • Immunology
  • Endocrinology

Background:

  • Accurate measurement of intact human proinsulin is crucial for diagnosing and managing metabolic disorders.
  • Existing assays may lack specificity or sensitivity for intact proinsulin.
  • Proinsulin levels are altered in conditions like diabetes and insulinoma.

Purpose of the Study:

  • To develop and validate a sensitive two-site sandwich enzyme-linked immunosorbent assay (ELISA) for quantifying intact human proinsulin.
  • To assess the assay's performance characteristics, including specificity, sensitivity, and precision.
  • To evaluate proinsulin concentrations in healthy individuals and patients with specific endocrine conditions.

Main Methods:

  • Development of a two-site sandwich ELISA utilizing monoclonal antibodies targeting specific proinsulin epitopes.

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  • Assessment of cross-reactivity with related peptides like insulin and C-peptide.
  • Determination of assay sensitivity, working range, intra- and interassay variability, and recovery.
  • Analysis of proinsulin levels in serum/plasma samples from healthy subjects and patients with non-insulin-dependent diabetes mellitus and insulinoma.

    • Main Results:

    • The developed ELISA demonstrated high sensitivity with a detection limit of 0.2 pmol/L and a working range of 0.2-100 pmol/L.
    • Negligible cross-reactivity was observed with insulin, C-peptide, and proinsulin conversion intermediates.
    • The assay exhibited excellent precision (intra-assay CV 2.4%, interassay CV 8.9%) and mean recovery of 103%.
    • Significantly elevated intact proinsulin levels were detected in patients with non-insulin-dependent diabetes mellitus (median 7.7 pmol/L) and insulinoma (median 153 pmol/L) compared to healthy subjects (median 2.7 pmol/L).

    Conclusions:

    • The described two-site sandwich ELISA is a sensitive and specific method for measuring intact human proinsulin in biological samples.
    • This assay can reliably differentiate proinsulin levels in healthy individuals versus those with specific metabolic and endocrine diseases.
    • The findings support the clinical utility of intact proinsulin measurement in the diagnosis and management of diabetes and insulinoma.