{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T00:06:25Z","timestamp":1771459585360,"version":"3.50.1"},"reference-count":23,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2025,5,1]],"date-time":"2025-05-01T00:00:00Z","timestamp":1746057600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Drug Testing and Analysis"],"published-print":{"date-parts":[[2025,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>The usage of ACE\u2010031 (Ramatercept), a dimeric fusion protein consisting of a human activin receptor IIB (ACVR2B) fragment linked to an Fc\u2010part of human IgG1, is banned according to chapter S4.3 of the \u201cWADA 2024 List of Prohibited Substances and Methods\u201d due to its potential performance enhancing properties. While ACE\u2010031 has not yet been pharmaceutically approved, it is sold as research chemical on the \u201cblack market\u201d (BM). The article presents a study on BM ACE\u2010031 products and its detection by gel\u2010electrophoresis and Western blotting. Of 14 tested products, only 12 contained an ACVR2B\u2010immunoreactive protein. Electrophoretic separation by SDS\u2010PAGE also showed that the 12 ACVR2B\u2010products contained many other proteins in addition to the main compound (ca. 58.4\u2009kDa). Further analyses by mass spectrometry and immunoblotting revealed that the 12 products contained the full\u2010length human activin receptor IIB instead of ACE\u2010031. The absence of an Fc\u2010fusion protein was further confirmed by treatment with IdeS protease, which was unable to cleave the BM products. In addition, it was demonstrated that the protocol we developed to detect luspatercept (another ACVR2B\u2010Fc fusion protein) in human serum could also be successfully applied for the detection of BM ACE\u2010031. Because administering black market products to human subjects was not ethically justifiable, a study was conducted with rats. In rat serum, BM ACE\u2010031 was detectable up to 48\u2009h post administration. However, due to the relatively high dose applied (10\u2009mg\/kg body weight) and possible differences in metabolism, the detection window may be different in humans.<\/jats:p>","DOI":"10.1002\/dta.3898","type":"journal-article","created":{"date-parts":[[2025,5,2]],"date-time":"2025-05-02T08:07:17Z","timestamp":1746173237000},"page":"1934-1946","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Gel Electrophoretic Detection of Black Market ACE\u2010031"],"prefix":"10.1002","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4256-8465","authenticated-orcid":false,"given":"Christian","family":"Reichel","sequence":"first","affiliation":[{"name":"Doping Control Laboratory Seibersdorf Seibersdorf Labor GmbH  Seibersdorf Austria"},{"name":"European Monitoring Center for Emerging Doping Agents German Sport University Cologne  Cologne Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Filip","sequence":"additional","affiliation":[{"name":"University of Veterinary Medicine Vienna  Vienna Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G\u00fcnter","family":"Gmeiner","sequence":"additional","affiliation":[{"name":"Doping Control Laboratory Seibersdorf Seibersdorf Labor GmbH  Seibersdorf Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1535-6451","authenticated-orcid":false,"given":"Mario","family":"Thevis","sequence":"additional","affiliation":[{"name":"European Monitoring Center for Emerging Doping Agents German Sport University Cologne  Cologne Germany"},{"name":"Institute of Biochemistry\/Center for Preventive Doping Research German Sport University Cologne  Cologne Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,5]]},"reference":[{"key":"e_1_2_8_2_1","volume-title":"International Nonproprietary Names for Pharmaceutical Substances (INN). 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