Development and validation of an immunoreceptor assay for simulect based on surface plasmon resonance.

Simulect is a chimeric human/mouse antibody directed against interleukin-2 (IL-2) receptor. A combined immuno- and receptor assay has been developed and validated to characterize the production of Simulect batches. This assay is based on surface plasmon resonance (SPR) technology. In each experiment two successive interactions were monitored: the direct binding of Simulect to an anti-human IgG antibody, followed by the direct binding of IL-2-soluble receptor to the preformed anti-human IgG antibody/Simulect complex. Based on the first interaction a direct immunoassay for Simulect was optimized and validated. Based on the second interaction a direct receptor assay for Simulect biological activity was optimized and validated. The assays were validated by performing three independent assays on 3 different days. The intra- and interday variations of the immunoassay (expressed as % CV) were, respectively, 1.7 and 1.6%. The overall accuracy for the immunoassay was 98.5% +/- 1. The intra- and interday variations of the receptor assay (% CV) were, respectively, 1.6 and 3.7%. The overall accuracy of the receptor assay was 100% +/- 2. Four batches of Simulect were compared to a reference batch. The results did not show significant differences for the immunoreactivity. However, the results of the receptor assay showed accuracies which were apparently higher than 100%. This was explained by a slight degradation of the reference batch after few years of storage. These results demonstrate the advantage of this method combining evaluation of the immunological and biological integrity of the drug and a high reproducibility in accuracy and precision of the biosensor-based technology.