Analytical validation of a homogeneous immunoassay for determination of mycophenolic acid in human plasma.

Mycophenolic acid (MPA) is an immunosuppression agent for the prophylaxis of organ rejection in patients receiving allogeneic transplants. The drug is administered based in 2 formulations, mycophenolate mofetil (MMF) and enteric-coated mycophenolate sodium (EC-MPS). MPA acts by specific, reversible, uncompetitive inhibition of inosine monophosphate dehydrogenase (IMPDH) and thus blocks the proliferation of both T- and B-activated lymphocytes. Therapeutic drug monitoring (TDM) constitutes an important part of immunosuppressive treatment because of the demonstrated significant intraindividual and interindividual variability of its pharmacokinetic behavior. TDM is required to optimize immunosuppressive efficacy. We present the analytical validation of a homogeneous particle-enhanced turbidimetric inhibition immunoassay (PETINIA) technique for determination of MPA in human plasma, and compare with a homogeneous enzyme immunoassay technique (EMIT; reference method), both methods adapted on a Dimension analyzer (Siemens). We examined 50 human plasma samples from kidney transplant recipients treated with MMF or EC-MPA, which were analyzed simultaneously by both methods. The interassay precision was 5.95% at a concentration of 1.0 µg/mL, 3.47% at 7.5 µg/mL, and 3.75% at 12.0 µg/mL. The bias of PETINIA-MPA for each of the 3 quality control sample was <3.0%. Least squares linear regression yielded an r-value of 0.994 with the following linear regression equation: PETINIA = 0.939 * EMIT - 0.063. Bland-Altman comparison presented a mean negative difference of -0.312 µg/mL (standard deviation [SD], 0.441), namely, -7.6% for PETINIA-MPA. The PETINIA assay for monitoring MPA concentrations is an acceptable method for routine clinical use, with interassay imprecision (% coefficient of variation) ranging from 5.9% to 3.7% below and above the therapeutic concentration range, respectively. In conclusion, MPA-EMIT and PETINIA-MPA methods on Dimension analyzer have a good correlation (r = 0.994), but PETINIA-MPA method demonstrates a negative average difference of -7.6% in comparison with EMIT-MPA method.

Evaluation of a fully automated method for the determination of sirolimus.

BACKGROUND: Sirolimus (SRL) is a macrocyclic lactone, indicated for prevention of organ rejection after kidney transplantation. Therapeutic drug monitoring of this agent constitutes an important part to immunosuppressive treatment because of its narrow window of therapeutic efficacy. Routine methods include manual pretreatment of samples. The aim of this study was to evaluate the performance characteristics of an automated immunoassay that does not require manual pretreatment to quantify SRL in whole blood on the Dimension analyzer. METHODS: We examined 50 whole blood samples collected routinely from kidney transplant patients treated with SRL. The samples were analyzed simultaneously by an immunoassay on an IMx analyzer (reference method), which requires manual pretreatment step versus a totally automated immunoassay on the Dimension analyzer, which does not require this pretreatment. RESULTS: The Dimension SRL assay had a functional sensitivity of ≤2.4 ng/mL. Total imprecision was 15.6% at a concentration of 2.8 ng/mL; 10% at 7.9 ng/mL; and 5.2% at 18.4 ng/mL. Least-squares linear regression analysis yielded an r-value of 0.973 with the following equation: SRL-D=1.204*SRL-IMx-0.251. Bland-Altman comparison showed a mean positive difference of 1.38 ng/mL (95% confidence interval, -1.10 to 3.82), namely, 17.2% for SRL Dimension. The Dimension assay to monitor SRL concentrations was an acceptable method for routine clinical use, with total assay imprecision (%CV) ranging from 10.0% to 5.2% within and above the therapeutic concentration range, respectively. CONCLUSION: SRL IMx and Sirolimus Dimension methods show a good correlation (r=0.973), but the SRL Dimension method demonstrated a positive average difference of 17.2% compared with the IMx method. The Dimension assay to monitor whole blood SRL concentration does not require a manual pretreatment step, reducing turnaround time and making this method an attractive alternative for SRL analysis.