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Related Experiment Videos

A C3 convertase assay for nephritic factor functional activity.

E Jelezarova1, M Schlumberger, S Sadallah

  • 1Institute of Biochemistry, Swiss Federal Institute of Technology, ETH-Zentrum, CH-8092, Zurich, Switzerland.

Journal of Immunological Methods
|April 9, 2001
PubMed
Summary
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C3 nephritic factor (C3NeF) stabilizes the C3 convertase, driving alternative pathway activation and C3 depletion. A new solid-phase assay quantifies this stabilizing activity for C3NeF IgG.

Area of Science:

  • Immunology
  • Complement System Biology

Background:

  • C3 nephritic factor (C3NeF) is an autoantibody targeting the C3 convertase.
  • C3NeF stabilizes the C3 convertase, leading to continuous alternative pathway activation and C3 depletion.
  • NeF is associated with membranoproliferative glomerulonephritis and partial lipodystrophy.

Purpose of the Study:

  • To develop a reproducible solid-phase assay for quantifying the functional activity of purified C3NeF IgG.
  • To characterize the stabilizing activity of C3NeF IgG on C3 convertase decay.

Main Methods:

  • Generated C3 convertase on immobilized C3b or C3b(2)-IgG complexes.
  • Assessed C3 convertase decay in the presence of normal IgG or NeF IgG, with or without Factor H.
  • Measured residual convertase activity using radiolabeled C3 and a capture system.

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Main Results:

  • Developed a solid-phase assay to measure C3NeF IgG functional activity.
  • Demonstrated dose-dependent stabilizing activity of NeF IgG on C3 convertase decay, even with Factor H present.
  • Observed similar NeF IgG stabilizing activity using both C3b and C3b(2)-IgG complexes.

Conclusions:

  • The developed solid-phase assay provides reproducible data on C3NeF IgG functional activity.
  • C3NeF IgG stabilizes the C3 convertase, confirming its role in alternative pathway dysregulation.
  • The assay is applicable to purified C3NeF IgG, including that isolated from urine.