Volume 9 Issue 1
Feb.  2019
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Rintaro Sogawa, Tetsuya Saita, Yuta Yamamoto, Sakiko Kimura, Yutaka Narisawa, Shinya Kimura, Masashi Shin. Development of a competitive enzyme-linked immunosorbent assay for therapeutic drug monitoring of afatinib[J]. Journal of Pharmaceutical Analysis, 2019, 9(1): 49-54.
Citation: Rintaro Sogawa, Tetsuya Saita, Yuta Yamamoto, Sakiko Kimura, Yutaka Narisawa, Shinya Kimura, Masashi Shin. Development of a competitive enzyme-linked immunosorbent assay for therapeutic drug monitoring of afatinib[J]. Journal of Pharmaceutical Analysis, 2019, 9(1): 49-54.

Development of a competitive enzyme-linked immunosorbent assay for therapeutic drug monitoring of afatinib

  • Publish Date: Feb. 10, 2019
  • Afatinib is an oral tyrosine kinase inhibitor (TKI) approved for treating advanced non-small cell lung cancer. It is necessary to develop a simple quantification method for TKIs in order to facilitate therapeutic drug monitoring (TDM) in clinical settings. This study sought to develop a simple and sensitive com-petitive enzyme-linked immunosorbent assay (ELISA) to quantify afatinib in plasma for routine phar-macokinetic applications. An anti-afatinib antibody was obtained using (S)-N-4-(3-chloro-4-fluor-ophenyl)-7-(tetrahydrofuran-3-yloxy)-quinazoline-4,6-diamine (CTQD), which has the same sub-structure as afatinib, as a hapten. Enzyme labeling of afatinib with horseradish peroxidase was similarly performed using CTQD. A simple competitive ELISA for afatinib was developed based on the principle of direct competition between afatinib and the enzyme marker for the anti-afatinib antibody, which had been immobilized on the plastic surface of a microtiter plate. Plasma afatinib concentrations below the limit of quantification of 30 pg/mL were reproducibly measurable. Also, the values of plasma afatinib levels measured from 20 patients were comparable with those measured by high-performance liquid chromatography, and there was a strong correlation between the values determined by both methods (Y = 0.976X – 0.207, r = 0.975). As indicated by its specificity and sensitivity, this newly developed ELISA for afatinib is an important tool for TDM and studies of the pharmacokinetics of afatinib.
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