Detection of Sotatercept (ACE‐011) in human serum by SAR‐PAGE and western single blotting |
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| Authors: | Christian Reichel Letizia Farmer Günter Gmeiner Katja Walpurgis Mario Thevis |
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| Institution: | 1. Doping Control Laboratory Seibersdorf, Seibersdorf Labor GmbH, Seibersdorf, Austria;2. Institute of Biochemistry/Centre for Preventive Doping Research, German Sport University Cologne, Germany;3. European Monitoring Centre for Emerging Doping Agents, German Sport University Cologne, Germany |
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| Abstract: | A method for the detection of Sotatercept (ACE‐011, ACVR2A‐Fc) in human serum is presented. The method is a modification of a recently published protocol for Luspatercept (ACE‐536, ACVR2B‐Fc), another erythropoiesis stimulating fusion protein. Out of 27 tested antibodies against either the extracellular domain of ACVR2A or the full‐length protein, only 4 antibodies bound strongly enough to Sotatercept for usage with immunoprecipitation followed by SAR‐PAGE and Western single blotting. The adapted protocol allows the detection of 0.1 ng/mL Sotatercept in just 50 μL human serum. None of the 3 commercial ACVR2‐ELISAs was able to detect Sotatercept and the 2 tested surrogate proteins, even in the μg/mL range. As for Luspatercept, only IPG‐IEF/2D‐PAGE generated discrete isoforms. Due to the long serum half‐life, the SAR‐PAGE method will be able to detect Sotatercept for several weeks and will be very useful in doping testing. |
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| Keywords: | activin receptor ACVR2A‐Fc doping control SAR‐PAGE Sotatercept |
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