Research on the standardization of the assays for anticardiolipin antibodies.

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Anticardiolipin antibodies (aCL) which recognize epitope on β2-glycoprotein I (β2-GPI) is a marker of antiphospholipid syndrome (APS), and considered to be pathogenic. However, the lack of a proper international standard for measurement of aCL and anti-β2-GPI antibodies make it difficult to compare data from different laboratories. To standardize the assay for aCLs and anti-β2-GPI antibodies is important for clinical investigation of APS and for studies on pathophysiology of antiphospholipid syndrome.We already established EY2C9, IgM monoclonal aCL derived from an APS patient, which is suitable for a standard of IgM aCL and anti-β2-GPI assays, but did not have a human IgG monoclonal aCL.Therefore, plasmids containing variable region genes of WBCAL-1, which is a monoclonal aCL derived from an APS prone mouse, and human γ1 and κ constant region genes were made. The constructs were transfected to P3U1, a mouse myeloma cell line and tarnsfected cells which secreted HCAL, a chimeric antibody, were cloned.EY2C9 and HCALA had the similar characteristics as aCL detected in sera from APS patients and high avidity to 2-GPI.Those antibodies worked well in most of the representative laboratories studying aCLs and ant-β2-glycoprotein I antibodies in the world, and are approved as standards for assay for aCLs by NIH and FDA.Cell line which secreting HCALA, a chimeric antibody consisting of variable regions of WBCAL-1 and human α1 and κ constant regions, was established in the same methods. HCALA recognized β2-GPI bound to CL or to oxygenated polystyrene surface. This binding profile is similar to that of aCL in sera from APS patients. The avidity of HCALA was high enough for a standard of aCL and anti-β2-GPI antibody assays.