Gel Electrophoretic Detection of Black Market ACE‐031

Title (eng)
Gel Electrophoretic Detection of Black Market ACE‐031
Author
Christian Reichel
Doping Control Laboratory Seibersdorf, Seibersdorf Labor GmbH, Seibersdorf, Austria
Author
Thomas Filip
Author
Günter Gmeiner
Doping Control Laboratory Seibersdorf, Seibersdorf Labor GmbH, Seibersdorf, Austria
Author
Mario Thevis
European Monitoring Center for Emerging Doping Agents, German Sport University Cologne, Cologne, Germany
Abstract (eng)
The usage of ACE-031 (Ramatercept), a dimeric fusion protein consisting of a human activin receptor IIB (ACVR2B) fragment linked to an Fc-part of human IgG1, is banned according to chapter S4.3 of the “WADA 2024 List of Prohibited Substances and Methods” due to its potential performance enhancing properties. While ACE-031 has not yet been pharmaceutically approved, it is sold as research chemical on the “black market” (BM). The article presents a study on BM ACE-031 products and its detection by gel-electrophoresis and Western blotting. Of 14 tested products, only 12 contained an ACVR2B-immunoreactive protein. Electrophoretic separation by SDS-PAGE also showed that the 12 ACVR2B-products contained many other proteins in addition to the main compound (ca. 58.4 kDa). Further analyses by mass spectrometry and immunoblotting revealed that the 12 products contained the full-length human activin receptor IIB instead of ACE-031. The absence of an Fc-fusion 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-Fc fusion protein) in human serum could also be successfully applied for the detection of BM ACE-031. Because administering black market products to human subjects was not ethically justifiable, a study was conducted with rats. In rat serum, BM ACE-031 was detectable up to 48 h post administration. However, due to the relatively high dose applied (10 mg/kg body weight) and possible differences in metabolism, the detection window may be different in humans.
Keywords (eng)
ACE-031Black MarketDopingMyostatinSDS-PAGE
Type (eng)
journal article
Language
[eng]
Persistent identifier
https://phaidra.vetmeduni.ac.at/o:4995
DOI
10.1002/dta.3898
Is in series
Title (eng)
Drug Testing and Analysis
Volume
17
Issue
10
ISSN
1942-7603
Issued
2025
Number of pages
13
From page
1934
To page
1946
Publication
Wiley
Version type (eng)
version of record (published version)
Date issued
2025
Access rights (eng)
open access
License
CC BY-NC-ND 4.0 International
Rights statement (eng)
© 2025 The Author(s)