In this study we sought to assess (1) the diagnostic value of a combined search for anti–β 2 -glycoprotein (aβ 2 -GPIs) and anticardiolipin antibodies (aCLs) in primary (APS I) and secondary (APS II) antiphospholipid syndrome and (2) the influence of the β 2 -GPI preparation in the ELISA's results. aβ 2 -GPI and aCL concentrations were assessed in 70 patients with APS and compared with those in 65 patients with systemic lupus erythematosus (SLE) without clinical features of APS. In APS patients (38 with APS I, 32 with APS II), the diagnosis had to have been made at least 3 years earlier; in subjects with SLE, the diagnosis had to have been made at least 5 years earlier. All serum samples were tested for aβ 2 -GPI with the use of an in-house ELISA with an aβ 2 -GPI preparation from human plasma. Samples negative for aβ 2 -GPI were controlled with 2 additional β 2 -GPI preparations, 1 from human serum and 1 from bovine serum. In APS, aβ 2 -GPIs were more frequent than in SLE (76% and 15%, respectively; P < .0001), mainly with IgG isotype and with significantly higher levels than those found in SLE. The specificity for APS was 92% for IgG aβ 2 -GPIs and 68% for IgG aCLs. The highest association with APS was found for the combination of the 2 markers (odds ratio 29; 95% confidence interval 10–76; P < .0001). Among the APS patients, 6 were positive for aCL only and remained negative regardless of which β 2 -GPI preparation was used; 1 patient was aCL-negative and only positive with human β 2 -GPI. These data emphasize the heterogeneity of the APS immunologic profile and the diagnostic possibilities of both antibodies.