Key result
The avidity of antibodies specifically bound to cardiolipin-coated wells was significantly higher than those bound non-specifically to cardiolipin-free wells.
Differentiating higher-avidity anticardiolipin antibodies from lower-avidity polyspecific antibodies using urea in ELISA may improve assay specificity and reduce background signal.
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May enhance anticardiolipin ELISA specificity; hypothesis-generating and requires clinical validation before practice change.
Avidity is an important feature of antibodies associated with their pathogenic effects. Anticardiolipin antibodies (aCLs) are the most commonly examined antiphospholipid antibodies, however, their avidity has been investigated only marginally. The aim of the study was to compare the avidity of the antibodies specifically bound to the cardiolipin-coated microtitrate wells with those bound to the wells bearing no cardiolipin in various conditions. We analysed 22 serum samples with high, medium and low levels of aCL IgG. The avidity of aCL IgG was determined in serially diluted sera by the ELISA method in the presence of increasing urea concentrations (from 2 to 8 mol/L) as a chaotropic agent. The serum dilution 1:50 and 1:100 and the concentrations of urea 6 mol/L and 8 mol/L seemed to be suitable for the determination of aCL avidity. The avidity of antibodies specifically bound to the cardiolipin and those bound to the cardiolipin-free wells significantly differed in their avidity. The simultaneous determination of higher-avidity aCL and lower-avidity polyspecific antibodies, whose presence complicates the ELISA methods by an increase of the background signal, might limit the shortcomings of some ELISA methods.
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Fialová et al. (2014) studied Antiphospholipid syndrome / Autoimmune and obstetric disorders (n=22). Cardiolipin-coated ELISA wells vs. Cardiolipin-free ELISA wells was evaluated on Avidity of IgG antibodies (Avidity Index) after exposure to urea (p=<0.005). The avidity of antibodies specifically bound to cardiolipin-coated wells was significantly higher than those bound non-specifically to cardiolipin-free wells.
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