Strategies for Effective Development of Ultra-Sensitive LC–MS/MS Assays: Application to a Novel STING Agonist

Abstract

Background: The continual need for the development and validation of ultra-sensitive (low pg/ml) LC–MS/MS assays in the pharmaceutical industry is largely driven by the ultra-low analyte exposure or very low sample volume. Methodology: Strategies and systematic approaches for sensitivity enhancement are provided which cover all aspects of a LC–MS/MS bioanalysis. A case study where such strategies were applied for the validation of a 5.0 pg/ml assay for a STING agonist is discussed. Conclusion: Analytical protocols were developed to extract analytes from large volume of plasma samples (600 and 400 μl) with high throughput. The guidance provided in this publication can serve as a resource to influence LC–MS/MS method development activities.

Keywords:

• carryover
• LC–MS/MS
• LLQ
• nonspecific binding
• pharmacokinetic
• sensitivity enhancement

Financial & competing interests disclosure

This research was supported by GlaxoSmithKline (GSK) Research and Development. All authors are employees of GSK and may be eligible for GSK stock options or have stock ownership. The authors have no other 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 apart from those disclosed.

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.