Related Experiment Video
Updated: Sep 12, 2025
Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells
Published on: January 29, 2014
The hemolytic activity of ABO antibodies: Correlation between quantitative tests, complement-mediated hemolysis assay
Sidneia Sanches de Menezes Costa1,2, Beatriz Girardo1,2, Thamy Caroline Silva1
1UNIFESP - Blood Center of the Federal University of São Paulo, São Paul, Brazil.
Background:
ABO antibodies can activate complement and cause hemolysis, sometimes with ABO minor incompatible platelet or plasma transfusions. Donor low titres of anti-ABO are used to assess transfusion safety. However, the correlation between antibody titres (measured semi-quantitatively or quantitatively) and their ability to activate complement is unclear.
Objectives:
This study aimed to correlate ABO antibody (IgM/IgG) titres and the presence of IgG1/IgG3 subclasses with complement-mediated haemolysis using the CHUHE-P assay (Complement Hemolysis Using Human Erythrocytes).
Methods:
Single-center blood group O donor samples were tested with semi-quantitative methods and classified in group 1 as low ABO titres if titre <100 and into group 2 as high ABO titre if titre >100. Testing included IgM/IgG antibody titration, the CHUHE-P assay, haemolysin test, and determination of IgG1/IgG3 subclasses.
Results:
Group 1 had 21 and Group 2 had 56 blood donors. In Group 2, CHUHE-P positivity was not associated with IgM or IgG anti-A/B titres or haemolysin test results. IgG1 and IgG3 subclasses of anti-A were significantly associated with positive CHUHE-P. Significant differences between groups included CHUHE-P positivity (p < 0.001), IgG titres (p < 0.01) for anti-A, and CHUHE-P positivity (p < 0.001), IgM titres (p = 0.04), and haemolysin test (p = 0.02) for anti-B. In Group 1, 33% of anti-A and 23.8% of anti-B had positive CHUHE-P.
Conclusions:
This study aimed to compare the ability of anti-ABO antibodies to cause complement-mediated hemolysis measured by the CHUHE-P test and the results of semi-quantitative/quantitative titration assays, the presence of IgG1 and/or IgG3, and hemolysin test results, where we highlighted an inconsistent association between the methods; the positivity for CHUHE-P was associated with the subclasses of antibodies IgG1 and IgG3.
Related Concept Videos
01:12The ABO Blood Group
Antigens in the ABO Blood Group System
Antigens are substances that can trigger an immune response, leading to the production of antibodies. In the ABO blood group system,...
01:10Blood Typing
Antigens are protein molecules that reside on the surface of red blood cells (RBCs). The ABO and Rh blood typing systems target...
02:45Blood Transfusion and Agglutination
History
The history of blood transfusion dates back to the 17th century, when early attempts were made in animals. In 1818 James Blundell, a British doctor, performed the first successful human blood transfusion. Later in 1900, Karl...
02:20Blood Types
ABO blood group
ABO antigens are glycoproteins encoded by genes present on...
01:26Antibody Actions
Neutralization
Antibodies can bind to pathogens, preventing them from infecting host cells. This process...
01:15Blood Transfusion
Blood Transfusion Overview
A blood transfusion is a medical procedure used to replace blood lost due to injury, surgery, or to treat conditions such as anemia or cancer. During a transfusion, donor blood is...