Novel nephelometric assay for measurement of complement 3d

Ann Clin Biochem. 2002 Jan;39(Pt 1):34-8. doi: 10.1258/0004563021901702.

Abstract

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.

MeSH terms

  • Antigen-Antibody Reactions
  • Complement C3d / analysis*
  • Hemoglobins / analysis
  • Humans
  • Immunodiffusion / methods
  • Lipids / analysis
  • Nephelometry and Turbidimetry / methods*
  • Reference Standards
  • Reference Values

Substances

  • Hemoglobins
  • Lipids
  • Complement C3d