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Related Experiment Videos

Effect of complement binding on a solid-phase immunometric TSH assay.

K Käpyaho1, P Tanner, T Weber

  • 1Minerva Institute for Medical Research, Kauniainen, Finland.

Scandinavian Journal of Clinical and Laboratory Investigation
|May 1, 1989
PubMed
Summary
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Serum components can interfere with diagnostic tests by binding to antibodies on plastic beads. This complement binding can be blocked by specific immune complexes, improving assay reliability.

Area of Science:

  • Immunology
  • Biochemistry
  • Diagnostic Assays

Background:

  • Solid-phase immunometric assays utilize antibody-coated surfaces for detecting analytes.
  • Serum components can potentially interfere with these assays, affecting accuracy.
  • The role of complement system activation in antibody-based assays was not fully understood.

Purpose of the Study:

  • To investigate the mechanism of inhibition observed in serum-based TSH assays.
  • To identify the inhibitory factor in serum and its interaction with antibody-coated surfaces.
  • To determine if complement activation plays a role in assay interference.

Main Methods:

  • Monoclonal antibody-coated plastic beads were used for TSH binding assays.
  • Serum was incubated with beads, with and without EDTA, to assess inhibition.

Related Experiment Videos

  • Heat lability and abrogation by albumin-anti-albumin complexes were tested.
  • Binding of the first component of complement (C1q) to antibody-coated beads was measured.

    • Main Results:

    • Serum inhibited TSH binding to antibody-coated beads, an effect reversed by EDTA.
    • The inhibitory factor was heat labile and its effect was blocked by albumin-anti-albumin complexes.
    • Antibody-coated beads were shown to bind C1q, which was inhibited by albumin-anti-albumin complexes.

    Conclusions:

    • Monoclonal antibody-coated surfaces can bind complement.
    • Complement binding to the antibody surface can interfere with solid-phase immunometric assays.
    • Understanding and mitigating complement interference is crucial for accurate diagnostic test results.