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Development of an Abbott ARCHITECT cyclosporine immunoassay without metabolite cross-reactivity
Elaine M Brate1, David M Finley, Jonathan Grote
1Abbott Diagnostics, Abbott Park, IL 60064, USA.
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.
