Multiplex Genotyping of CYP3A4, CYP3A5, CYP2C9 and CYP2C19 SNPs Using MALDI-TOF Mass Spectrometry

Abstract

Background: Pharmacogenetics is the study of genetic variations that cause alterations in drug level, drug response and adverse drug reactions. SNPs found in CYP450 genes have the greatest genetic influences on interindividual variability in drug bioavailability. The polymorphic nature of these genes may modulate several enzyme levels that affect individual responses to pharmacological treatment. Among them, CYP3A4, CYP3A5, CYP2C9 and CYP2C19 isoforms of CYP450 enzymes are involved in the metabolism of many commonly prescribed drugs. Aims: In this study, we would like to develop a CYP450 genotyping platform that could lead a complete definition of a patient‘s metabolic genotype in order to improve the clinical outcome of some drug treatments. Materials & methods: We used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (Sequenom^®) to develop a SNP genotyping method. Results: This MALDI-TOF-based multiplexing system allows the simultaneous and efficient genotyping of a set of CYP450 gene polymorphisms. Conclusion: The multiple CYP450 gene testing achieved with this application can be used to develop diagnostic tests to predict drug responses and clinical outcomes.

KEYWORDS::

• CYP2C9
• CYP2C19
• CYP3A4
• CYP3A5
• CYP450
• drug treatment
• individualized therapy
• mass spectrometry
• pharmacogenetics

Acknowledgements

The authors gratefully acknowledge for their assistance, useful discussions and comments provided: Dr Paolo Lazzari, PharmaNess Head of Research and Development, Dr Luigi Pira, PharmaNess Head of General Pharmacology and Dr Enrico Sorisio, PharmaNess Managing Director.

Financial & competing interests disclosure

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.