Chemiluminescence immunoassay of cyclosporine in whole blood

Abstract

We describe a chemiluminescent immunoassay (CLI) for measuring cyclosporine in whole blood. Its sensitivity and accuracy are comparable with those of an RIA method that makes use of the same specific monoclonal antibody. The comparison with the RIA method was excellent: y(RIA) = x(CLI) + 11.24 (r = 0.99). In our procedure the samples are incubated with cyclosporin C-hemisuccinate-aminobutyl-N-ethylisoluminol, antibody, and paramagnetic particles coated with second antibody. After magnetic separation and washing, the samples are incubated with 200 μL of NaOH (2 mol/L) at 60 °C for 30 min. The chemiluminescence generated by automated serial injections of solutions of microperoxidase and dilute (2 mL/L) H2O2 is measured for 5 s. The data are processed by using a spline fit of log B/Bo log conversion. This method is easy to perform and avoids the hazards and costs associated with isotopic waste disposal. The label is stable for at least three years.