Related Experiment Video
Updated: May 3, 2026
06:29
Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells
Published on: January 29, 2014
29.8K
Methods for quantitative detection of antibody-induced complement activation on red blood cells
Elisabeth M Meulenbroek1, Diana Wouters1, Sacha Zeerleder1,2
1Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Center, University of Amsterdam.
Journal of Visualized Experiments : Jove
|February 12, 2014
Summary
New quantitative assays detect complement activation on red blood cells (RBCs) caused by antibodies. These reproducible methods aid in studying antibody-induced complement activation and evaluating complement inhibitors for conditions like autoimmune hemolytic anemia (AIHA).
Area of Science:
- Immunology
- Hematology
- Biochemistry
Background:
- Antibodies binding to red blood cells (RBCs) can trigger complement activation, leading to hemolysis (RBC destruction) via intravascular or extravascular pathways.
- This complement activation, particularly intravascular hemolysis, poses significant health risks and can be fatal, as seen in autoimmune hemolytic anemia (AIHA).
- Current diagnostic methods like the Coombs test are often non-quantitative, limiting their utility in assessing the efficacy of complement inhibitors.
Purpose of the Study:
- To develop and present novel, quantitative assays for assessing complement activation on RBCs induced by patient antibodies.
- To provide tools for accurately measuring complement deposition and hemolysis, crucial for evaluating complement-targeted therapies.
Main Methods:
- Quantification of C3 and C4 deposition on RBCs using flow cytometry (FACS) with fluorescently labeled antibodies.
- Measurement of complement-mediated RBC lysis using a quantitative hemolytic assay with spectrophotometric detection of released hemoglobin.
Main Results:
- Developed two reproducible, quantitative assays to measure antibody-induced complement activation on RBCs.
- These assays accurately detect complement deposition (C3, C4) and hemolysis, offering a significant improvement over existing non-quantitative methods.
Conclusions:
- The new FACS and hemolytic assays provide robust, quantitative measures of complement activation on RBCs.
- These assays are essential for research into antibody-mediated RBC destruction and for the clinical evaluation of complement inhibitors in conditions like AIHA.
