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  • Title: Development of an Abbott ARCHITECT cyclosporine immunoassay without metabolite cross-reactivity.
    Author: Brate EM, Finley DM, Grote J, Holets-McCormack S, Ozaeta PF, Pacenti D, Peart JE, Piktel RE, Ramsay CS, Rupprecht KR, Saldana SC, Spring TG, Tetin SY, Trudeau BC, Wang P, Xie H.
    Journal: Clin Biochem; 2010 Sep; 43(13-14):1152-7. PubMed ID: 20599875.
    Abstract:
    OBJECTIVE: We investigated the mechanism by which the ARCHITECT cyclosporine (CsA) chemiluminescent microparticle immunoassay (CMIA) eliminates cross-reactivity to CsA metabolites AM1 and AM9, despite its use of a monoclonal antibody which shows cross-reactivity in fluorescence polarization immunoassays. DESIGN AND METHODS: The CMIA was accomplished by incubating an extracted blood sample with magnetic microparticles coated with a very low amount of anti-CsA antibody. After a wash step the microparticles were incubated with a chemiluminescent CsA tracer, followed by a second wash step and measurement of chemiluminescence. The reagent concentrations of salt and detergent were optimized to maximize CsA binding and minimize metabolite interference. RESULTS: Elimination of CsA metabolite cross-reactivity was shown using purified metabolites and blood samples containing native CsA metabolites. The CMIA demonstrated precision and sensitivity acceptable for use in a clinical setting. CONCLUSION: We conclude that it is possible to eliminate CsA metabolite immuno-cross-reactivity by careful assay design.
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