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Updated: Aug 16, 2026
Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells
Published on: January 29, 2014
Novel nephelometric assay for measurement of complement 3d
Xavier Bossuyt1, Lutgarde Sneyers, Godelieve Mariën
1Laboratory Medicine, Immunology, University Hospitals Leuven, Belgium. xavier.bossuyt@uz.kuleuven.ac.be
Background:
This report describes a nephelometric assay for measuring complement 3d (C3d).
Methods:
C3d was separated from C3 and the other C3 split products by polyethylene glycol precipitation. The supernatant containing C3d was then measured by rate nephelometry in a Beckman Immage rate nephelometer using a specific antibody. Calibration was performed with dilutions of a standard, which was obtained by incubating a serum pool at 37 degrees C for 7 days. This incubation step allowed the activation and breakdown of C3 into C3d.
Results:
The assay was linear across the normal and pathological range. Precision studies showed within-run and between-run coefficients of variation of < 5% and < 6%, respectively. The nephelometric results are comparable with those obtained by radial immunodiffusion. Neither haemoglobin nor lipids interfere with the assay.
