SUMMARYWe examined the role of autoantibodies to b 2 -GPI and prothrombin (PT) in the inhibition of annexin V binding to cardiolipin (CL) and the association with clinical manifestations of the anti-phospholipid syndrome (APS). Plasma samples from 59 patients with anti-phospholipid (aPL) antibodies were studied. Affinity purification of total IgG and IgG anti-û 2 -GPI antibodies was performed using staphylococcal protein A and phospholipid liposomes. Annexin V binding to CL was significantly inhibited by 31/59 (53%) aPL 1 plasma samples. There was a significant association between annexin V inhibition and elevated levels of IgG anti-cardiolipin (aCL) (r 20´62; P , 0´001), IgG anti-û 2 -GPI (r 20´67; P , 0´001) and a weaker association with lupus anti-coagulant (r 20´27; P 0´05). There was no association with other isotypes of aCL and anti-û 2 -GPI or with anti-PT of any isotype. In patients with clinical manifestations of the APS there were higher levels of IgG aCL (median (range) Z score): 10´0 (0± 17´6) versus 5´0 (0±16´1); P 0´03), IgG anti-û 2 -GPI (4´5 (0±11´3) versus 0´9 (0±9´7); P 0´02) and greater inhibition of annexin V binding to CL (23´4 (211´4±0´6) versus 21´1 (210´8±1´2); P 0´22). Odds ratios for the laboratory assays and the presence of clinical manifestations of the APS varied between 0´38 and 4´16, with the highest values for IgG aCL (4´16), IgG anti-û 2 -GPI (3´28) and annexin V inhibition (2´85). Additional experiments with affinity-purified IgG antibodies indicated that inhibition of annexin V binding was dependent upon the concentration of û 2 -GPI and anti-û 2 -GPI antibodies. These results indicate that inhibition of annexin V binding to procoagulant phospholipid surfaces is dependent upon anti-û 2 -GPI antibodies and suggest a role for annexin V in the pathogenesis of the APS.