OBJECTIVES: The Certified Reference Material (CRM) ERM^®-DA477/IFCC is a new polyclonal IgG anti-beta2-glycoprotein I (anti-β2GPI) material for the harmonization of the laboratory diagnosis of antiphospholipid syndrome (APS). We evaluated CRM's ability to represent the heterogeneity of APS patient anti-β2GPI antibodies and to calibrate IgG anti-β2GPI methods.

METHODS: We characterized CRM for its reactivity against domain-1, using the QUANTA Flash^® β2GPI-domain-1 assay, and against domains-4-5 of β2GPI, and single-domain-deleted β2GPI molecules using in-house ELISAs. We used QUANTA Lite^® ELISA, QUANTA Flash^® CLIA, and EliA™ FEIA methods to evaluate the CRM's anti-Cardiolipin (anti-CL) activity. Four anti-β2GPI IgG methods (in-house and QUANTA Lite^® ELISA, QUANTA Flash^® CLIA, and EliA™ FEIA) were also used to evaluate the CRM's calibration efficacy, alongside 133 clinical samples (CSs) and 99 controls.

RESULTS: The CRM showed high anti-domain-1 activity and no anti-domain-4-5 activity at the recommended assay dilution. The domain-dependent-β2GPI reactivity profiles were comparable with full-blown APS. There was acceptable dilution linearity for anti-CL assays with R² ranging from 0.957 to 0.997. For the four anti-β2GPI IgG assays, calibration with the CRM led to a good comparability of the average result of CSs for two of the assays. New cut-offs calculated from this work improved comparability in quantitative results between three of the assays: 85 % concordance with CRM compared to 66 % concordance with assay-specific-calibration.

CONCLUSIONS: The CRM is representative of patient anti-β2GPI/CL heterogeneity and should improve anti-β2GPI IgG method harmonization. However, the level of achievable method harmonization is affected by differences in the selectivity among the assays.