Development of an Abbott ARCHITECT cyclosporine immunoassay without metabolite cross-reactivity

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. The C...

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Published in:	Clinical biochemistry Vol. 43; no. 13; pp. 1152 - 1157
Main Authors:	Brate, Elaine M., Finley, David M., Grote, Jonathan, Holets-McCormack, Shelley, Ozaeta, Pan F., Pacenti, David, Peart, Joan E., Piktel, Ryan E., Ramsay, Carol S., Rupprecht, Kevin R., Saldana, Sylvia C., Spring, Thomas G., Tetin, Sergey Y., Trudeau, Beth C., Wang, Phil, Xie, Helen
Format:	Journal Article
Language:	English
Published:	Amsterdam Elsevier Inc 01-09-2010 Elsevier
Subjects:	Antibodies, Monoclonal Biological and medical sciences Chemiluminescence Cross Reactions Cross-reactivity Cyclosporine Cyclosporine - blood Cyclosporine - metabolism Fluorescence polarization Humans Immunoassay Immunoassay - methods Immunoassay - standards Immunomodulators Luminescent Measurements Medical sciences Metabolites Microparticle Monoclonal antibody Pharmacology. Drug treatments Sensitivity and Specificity Immunoassay Fluorescence polarization Metabolites Cross-reactivity Microparticle Chemiluminescence Cyclosporine Monoclonal antibody Human Calcineurin inhibitor Metabolite Interference Immunological method Optical polarization Cross reaction Ciclosporin Immunosuppressive agent
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Summary:	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. 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. 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. We conclude that it is possible to eliminate CsA metabolite immuno-cross-reactivity by careful assay design.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0009-9120 1873-2933
DOI:	10.1016/j.clinbiochem.2010.06.007