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A novel method using formamide for the elution of antibodies from erythrocytes
L Caruccio1, K Byrne, J Procter
1National Institutes of Health, Warren Grant Magnuson Clinical Center, Department of Transfusion Medicine, Bethesda Maryland, USA. lcaruccio@mail.cc.nih.gov
Background And Objectives:
Accurate identification of antibodies that sensitize red blood cells (RBCs) involves dissociating them from RBCs using an in vitro elution method that does not alter their antigen-binding properties, and analysis of the eluates against a panel of RBCs.
Materials And Methods:
A method was developed that allowed efficient RBC antibody elution. Human polyclonal anti-D was used to sensitize Rh-positive RBCs, and known antigen-antibody disruptive reagents were tested using these RBCs. The best reagent conditions were optimized. Eluates made were tested and compared to results obtained with a glycine-acid-based commercial elution kit to determine efficacy. Patient samples that were positive with direct antiglobulin tests (DATs), and in vitro commercial antisera-sensitized RBCs representing clinically significant antibodies, were used for evaluating the new method.
Results:
The formamide method was efficient at removing antibodies from RBCs. The patient samples with a positive DAT had antibodies recovered with the same specificity when compared to the acid-based technique. The length of preparation time was similar for both formamide and acid-based methods. Results of testing the eluates made from reagent RBCs sensitized with commercial antisera were distinct with antigen-positive and -negative erythrocytes.
Conclusions:
The formamide method compares well with acid techniques and may be an alternative choice of elution method.
