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Clinical Chemistry 38: 1307-1310, 1992;
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Clinical Chemistry, Vol 38, 1307-1310, Copyright © 1992 by American Association for Clinical Chemistry

Radioreceptor assay for quantifying FK-506 immunosuppressant in whole blood

JN Murthy, Y Chen, VS Warty, R Venkataramanan, JG Donnelly, A Zeevi and SJ Soldin
Department of Laboratory Medicine, Children's National Medical Center, Washington, DC 20010.

We describe a quantitative radioreceptor assay (RRA) for quantifying FK- 506 in whole blood. FK-506 extracted from whole blood with a cyclohexyl- sorbent column competes with [3H]dihydro-FK-506 for binding to a partially purified preparation of FK-506 binding protein (FK-BP). Free and protein-bound FK-506 are separated on LH 20 Sephadex chromatographic columns. We compared the results of this method with those of an enzyme immunoassay that uses a monoclonal antibody: r = 0.97, Sy/x = 0.039. Between-day precisions (CV) at FK-506 concentrations of 8 and 17 micrograms/L were 9.2% and 8.2%, respectively. Within-run precisions were 5.9%, 8.1%, and 9.4%, respectively, at 4, 8, and 15 micrograms/L. Analytical recovery, evaluated at 5, 10, 15, 20, and 25 micrograms/L for FK-506 added to whole blood, ranged from 98% to 103%. The assay can reliably quantify FK-506 blood concentrations between 1.0 and 25 micrograms/L.


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