Two kinds of plasmid expression vectors which expressed beta 2-glycoprotein 1 (beta 2GP1) and the fifth domain of beta 2-glycoprotein 1 (beta 2GP1-D5) were constructed respectively in this study. The antigenicity of recombinant beta 2GP1 (r beta 2GP1) and beta 2GP1-D5 (r beta 2GP1-D5) was identified by immunoblots using rabbit anti-beta 2GP1 antibodies, and the recombinant proteins were purified. Both anti-r beta 2GP1 and anti-beta 2GP1-D5 antibodies in 112 patients were detected by ELISA using r beta 2GP1 and r beta 2GP1-D5 as coating antigens. A significant statistical correlation (r = 0.667, P < 0.01) between the levels of anti-beta 2GP1 and anticardiolipin (ACL) antibodies was found. The presence of anti-r beta 2GP1 antibodies was associated with an increased frequency of history of thrombosis and/or recurrent abortion; hence anti-r beta 2GP1 assay provided better specificity than conventional ACL assay. Detection of anti-r beta 2GP1 antibodies may be of potential value in evaluating the risk of thrombosis and/or symptoms associated with other antiphospholipid syndromes (APS). The binding of anti-r beta 2GP1 from the sera of patients with APS to r beta 2GP1 was inhibited by r beta 2GP1-D5. Meanwhile, of 28 patients who had positive anti-r beta 2GP1 antibodies in sera, 27 (96.4%) had positive anti-r beta 2GP1-D1 antibodies. This indicated that the antigenic epitope of beta 2GP1 may be located in its fifth domain.