Laboratory Testing for Non-criteria Antiphospholipid Antibodies: Antibodies Toward the Domain I of Beta2-Glycoprotein I (aDI)

Abstract

Anti-β2GPI antibodies (aβ2GPI) are one of the laboratory criteria for antiphospholipid syndrome (APS), along with lupus anticoagulant (LA) and anticardiolipin antibodies (aCL). A subset of the aβ2GPI are the antibodies directed toward the domain I of the β2GPI (aDI). The aDI are regarded as non-criteria aPL and are among the most studied non-criteria aPL. Antibodies directed against a specific epitope in the domain I (G40-R43) of β2GPI were shown to be strongly correlated with thrombotic and obstetric events in APS. Many studies illustrated the pathogenic capacity of these antibodies, although with various results, depending on the assay used. The first studies were performed with an in-house ELISA with high specificity for aDI toward the G40-R43 epitope. More recently, a commercial chemiluminescence immunoassay for aDI IgG became obtainable for diagnostic laboratories. Although the added value of aDI on top of the criteria aPL is not clear, with opposing findings in literature, the assay might help in the diagnosis of APS, identifying the patients at risk since aDI are frequently present with high titers in triple-positive patients (positive for LA, aβ2GPI, and aCL). aDI can be used as a confirmatory test and is useful for proving the specificity of the aβ2GPI antibodies. In this chapter, the procedure for detecting these antibodies is outlined, using an automated chemiluminescence assay which can be used to determine the presence of IgG aDI in human samples. General guidelines that will facilitate optimal performance of the aDI assay are also provided.

Keywords: Antiphospholipid; Beta2-glycoprotein I; Chemiluminescence; Domain I; ELISA; Immunoassays; Protocol.