2. Academic Validation
  3. Development and validation of an ELISA with high sensitivity for therapeutic monitoring of afatinib

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

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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