Development and validation of an ELISA with high sensitivity for therapeutic monitoring of afatinib

  • Bioanalysis. 2018 Sep 1;10(18):1511-1523. doi: 10.4155/bio-2018-0095.
Mona M Al-Shehri  1 ,  Manal A El-Gendy  1 ,  Adel S El-Azab  1 ,  Mohammed A Hamidaddin  1  2 ,  Ibrahim A Darwish  1
Affiliations
  • 1. Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh 11451, Saudi Arabia.
  • 2. Department of Medicinal & Analytical Chemistry, Faculty of Pharmacy, Sana'a University, Sana'a, Yemen.
PMID: 30117333 DOI: 10.4155/bio-2018-0095
Abstract

Aim: To support the therapeutic drug monitoring of afatinib (AFT), an ELISA was required.

Results: A hapten for AFT was prepared and linked to each of BSA and KLH proteins by diazotization/coupling reaction. A polyclonal antibody recognizing AFT with high affinity (IC50 = 40 ng ml-1) was generated and used in the development of a competitive ELISA for quantitation of AFT in plasma samples. The assay limit of detection was 2 ng ml-1. The assay accuracy and precision were proved.

Conclusion: The assay is an appropriate alternative to the existing LC-MS/MS assays for AFT and it is anticipated to effectively contribute to the therapeutic drug monitoring of AFT in clinical settings.

Keywords
ELISA; afatinib; metastatic non-small-cell lung cancer; polyclonal antibody; therapeutic drug monitoring.
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