Universität zu Köln

Reichel, Christian, Gmeiner, Guenter and Thevis, Mario (2022). Electrophoretic detection of black market myostatin propeptide. Drug Test. Anal., 14 (11-12). S. 1812 - 1825. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

Myostatin propeptide is prohibited according to chapter S4 of the WADA 2022 List of Prohibited Substances and Methods. So far, no approved myostatin-propeptide pharmaceuticals are available. Nevertheless, myostatin-propeptides can be bought on the black market for research purposes. A study on black market myostatin propeptide products is presented as well as electrophoretic detection methods for serum and urine. Out of the 12 tested products, only nine actually contained the protein. Separation by SDS-PAGE revealed that the nine products were relatively impure and that the main compound had a much higher mass (approximately 54-55 kDa) than expected (approximately 33 kDa). Further analyses by mass spectrometry showed that the elevated molecular mass was due to the presence of a full length GST-tag on the propeptide. The developed detection method for serum is based on immunoprecipitation (IP) followed by SDS-PAGE and Western blotting. In total, three anti-myostatin propeptide antibodies were tested. All of them were well suited for either IP or immunoblotting. The final protocol applies a biotinylated polyclonal antibody, streptavidin-coated magnetic beads, and a monoclonal detection antibody. For a sample volume of 500 mu L serum, the detection limit of the method is approximately 2.5 ng/mL. The urine method applies a commercial ELISA for IP and performs with a limit of detection (LOD) of approximately 0.4 ng/mL. Furthermore, practically all currently available myostatin propeptide standards were also investigated. Due to the significant molecular mass difference of the black market products, an unambiguous differentiation from endogenous myostatin propeptide is possible.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Reichel, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Gmeiner, Guenter UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-665686
DOI:	10.1002/dta.3398
Journal or Publication Title:	Drug Test. Anal.
Volume:	14
Number:	11-12
Page Range:	S. 1812 - 1825
Date:	2022
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1942-7611
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language SKELETAL-MUSCLE; EXPRESSION; PROTEINS; MASS; ERYTHROPOIETINS; RECOMBINANT; MODEL; BONE; GENE; PAGE Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/66568