Functional analysis of the classical, alternative, and MBL pathways of the complement system: standardization and validation of a simple ELISA

J Immunol Methods. 2005 Jan;296(1-2):187-98. doi: 10.1016/j.jim.2004.11.016. Epub 2004 Dec 15.


Primary defence against invading microorganisms depends on a functional innate immune system and the complement system plays a major role in such immunity. Deficiencies in one of the components of the complement system can cause severe and recurrent infections, systemic diseases, such as systemic lupus erythematosus (SLE) and renal disease. Screening for complement deficiencies in the classical or alternative complement pathways has mainly been performed by haemolytic assays. Here, we describe a simple ELISA-based format for the evaluation of three pathways of complement activation. The assays are based on specific coatings for each pathway in combination with specific buffer systems. We have standardized these assays and defined cut off values to detect complement deficiencies at the different levels of the complement system. The results demonstrate the value of these ELISA-based procedures for the functional assessment of complement deficiencies in clinical practice. The assay is now available commercially in kit form.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Complement Activation / immunology*
  • Complement Pathway, Alternative
  • Complement Pathway, Classical
  • Complement Pathway, Mannose-Binding Lectin
  • Complement System Proteins / analysis
  • Complement System Proteins / deficiency*
  • Complement System Proteins / immunology
  • Enzyme-Linked Immunosorbent Assay / standards*
  • Humans
  • Lupus Erythematosus, Systemic / diagnosis
  • Lupus Erythematosus, Systemic / immunology
  • Mannose-Binding Lectin / blood
  • Mannose-Binding Lectin / immunology
  • Reagent Kits, Diagnostic*


  • Mannose-Binding Lectin
  • Reagent Kits, Diagnostic
  • Complement System Proteins