LC–MS/MS Strategies for Therapeutic Antibodies and Investigation Into The Quantitative Impact of Antidrug-Antibodies

Abstract

Aim: We aimed to establish novel, high-throughput LC–MS/MS strategies for quantification of monoclonal antibodies in human serum and examine the potential impact of antidrug antibodies. Methodology: We present two strategies using a thermally stable immobilized trypsin. The first strategy uses whole serum digestion and the second introduces Protein G enrichment to improve the selectivity. The impact of anti-trastuzumab antibodies on the methods was tested. Conclusion: Whole serum digestion has been validated for trastuzumab (LLOQ 0.25 µg/ml). Protein G enrichment has been validated for trastuzumab (LLOQ 0.1 µg/ml), bevacizumab (LLOQ 0.1 µg/ml) and adalimumab (LLOQ 0.25 µg/ml). We have shown the potential for anti-drug antibodies to impact on the quantification and we have subsequently established a strategy to overcome this impact where total quantification is desired.

Keywords::

• ADA (antidrug antibodies)
• adalimumab
• antibody–drug conjugates
• bevacizumab
• digestion
• LC–MS/MS
• monoclonal
• trastuzumab

Supplementary Data

Financial & competing interests disclosure

The authors are grateful to the Covance Scientific Council for financial backing of this research, and to Colin McKee of ADC Biotechnology (Denbighshire, UK) for supply of the trastuzumab-MMAE antibody–drug conjugate reference material. The authors are employees of Covance Laboratories Ltd. 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.