Research Article

Development of two complementary LC–HRMS methods for analyzing sotatercept in dried blood spots for doping controls

Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Cologne, Germany

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Katja Walpurgis

Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Cologne, Germany

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Andreas Thomas

Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Cologne, Germany

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Hans Geyer

Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Cologne, Germany

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Mario Thevis

*Author for correspondence: Tel.: +49 221 4982 7070; Fax: +49 221 4982 7071;

E-mail Address: m.thevis@biochem.dshs-koeln.de

Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Cologne, Germany

European Monitoring Center for Emerging Doping Agents (EuMoCEDA), Center for Preventive Doping Research, German Sport University, Cologne/Bonn, Germany

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Published Online:20 Jun 2019https://doi.org/10.4155/bio-2018-0313

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Abstract

Aim: sotatercept is a therapeutic Fc-fusion protein with erythropoiesis-stimulating activity. Due to a potential abuse of the drug by athletes in professional sports, a sensitive detection method is required. In sports drug testing, alternative matrices such as dried blood spots (DBS) are gaining increasing attention as they can provide several advantages over conventional matrices. Materials & methods: Herein, two complementary LC–high-resolution mass spectrometry (HRMS) detection methods for sotatercept from DBS, an initial testing procedure (ITP) and a confirmation procedure (CP) were developed and validated for the first time. Both methods comprise an ultrasonication-assisted extraction, affinity enrichment, proteolytic digestion and HRMS detection. Results & conclusion: For the multianalyte ITP, artificial samples fortified with sotatercept, luspatercept and bimagrumab, and authentic specimens containing bimagrumab were successfully analyzed as proof-of-concept. The validated detection methods for sotatercept are fit for purpose and the ITP was shown to be suitable for the detection of novel IgG-based pharmaceuticals in doping control DBS samples.

Keywords:

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Vol. 11, No. 10

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Received 7 December 2018

Accepted 21 March 2019

Published online 20 June 2019

Published in print May 2019

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Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: https://www.futuremedicine.com/doi/suppl/10.4155/bio-2018-0311

Acknowledgments

The authors thank Olivier Petricoul (Novartis, Switzerland) for providing the post-administration serum samples for proof-of-concept analyses.

Financial & competing interests disclosure

The study was conducted with support of the World Anti-Doping Agency (grant #ISF17A12MT), the Federal Ministry of the Interior of the Federal Republic of Germany (Berlin, Germany) and the Manfred-Donike Institute for Doping Analysis (Cologne, Germany). 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, and informed consent has been obtained from the participants involved.

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