Investigation of the crossreactivity of mycophenolic acid glucuronide metabolites and of mycophenolate mofetil in the Cedia MPA assay

Abstract

The immunosuppressant mycophenolic acid (MPA) used for solid organ transplantation is predominantly metabolized to a pharmacologically inactive phenolic glucuronide (MPAG) and, to a lesser extent, to the pharmacologically active acyl glucuronide (AcMPAG). The recently introduced CEDIA Mycophenolic Acid Assay from Microgenics has been reported to overestimate MPA in clinical samples and crossreactivity with AcMPAG has been suspected. A detailed investigation of the crossreactivity of AcMPAG and the prodrug mycophenolate mofetil (MMF) in the CEDIA assay is presented using pure substances. In addition, MPA concentrations in plasma were compared with a validated high-performance liquid chromatography-ultraviolet method. Plasma samples from kidney (KTx, n = 50), heart (HTx, n = 50), and liver (LTx, n = 50) transplant recipients were analyzed by the CEDIA (MPA) and a high-performance liquid chromatography-ultraviolet method (MMF, MPA, MPAG, AcMPAG). Crossreactivity of MMF (0.93-46.3 mg/L), MPAG (50-1000 mg/L), and AcMPAG (0.5-10 mg/L) was investigated using spiked drug-free plasma. Method comparison was performed using Bland & Altman and Passing & Bablok analysis. The method bias was correlated to AcMPAG concentrations using Spearman's rank correlation. Crossreactivity with AcMPAG and MMF was concentration-dependent and reached 215% and 143%, respectively. There was no crossreactivity with MPAG. The CEDIA assay showed a mean positive bias of 36.3% in patient samples. The mean bias was lowest with HTx samples (15%), 41.7% with KTx samples, and highest with LTx samples (52.3%). There was a positive correlation between the method bias and AcMPAG concentrations (r = 0.829; P < 0.001). No MMF was detected in patient samples. The CEDIA overestimates MPA concentrations on average by 36%. This bias is mainly the result of AcMPAG as previously observed with the EMIT MPA assay. It should be considered that the putative therapeutic range for MPA with the CEDIA assay will be higher than the range using high-performance liquid chromatography.