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

Mona M Al-Shehri; Manal A El-Gendy; Adel S El-Azab; Mohammed A Hamidaddin; Ibrahim A Darwish

doi:10.4155/bio-2018-0095

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. Epub 2018 Aug 17.

Authors

Mona M Al-Shehri¹, Manal A El-Gendy¹, Adel S El-Azab¹, Mohammed A Hamidaddin^{1

2}, Ibrahim A Darwish¹

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.

Publication types

Validation Study

MeSH terms

Afatinib / blood
Afatinib / pharmacology*
Afatinib / therapeutic use
Animals
Drug Monitoring / methods*
Enzyme-Linked Immunosorbent Assay / methods*
Rats

Substances

Afatinib