. 2019 Nov;11(11-12):1675-1697.
doi: 10.1002/dta.2741.
Affiliations
PMID: 31758732 DOI: 10.1002/dta.2741
Detection of black market follistatin 344
Christian Reichel et al. Drug Test Anal. 2019 Nov.
Erratum in
Detection of black market follistatin 344.
Reichel C, Gmeiner G, Thevis M. Reichel C, et al. Drug Test Anal. 2020 Oct;12(10):1522-1533. doi: 10.1002/dta.2882. Epub 2020 Sep 7. Drug Test Anal. 2020. PMID: 33460286 No abstract available.
Abstract
Follistatin, a myostatin-inhibiting protein, is prohibited according to chapter S4 of the "WADA 2019 List of Prohibited Substances and Methods". While currently no approved pharmaceutical formulations of follistatin are available, follistatin can be bought on the black market. Most of the products are labeled "follistatin 344" (FS344), a few "follistatin 315". A study on FS344 black market products was performed and an electrophoretic detection method for serum and urine developed. While only nine of the 17 tested products actually contained follistatin, in some of the others growth promoting peptides were found (e.g. MGF, GHRP-2). Surprisingly, all nine products contained His-tagged FS344 and a high degree of its oligomers. The detection method is based on immunomagnetic purification followed by SDS-PAGE and Western blotting with a monoclonal anti-His antibody. Alternatively, a monoclonal anti-follistatin antibody can be used. For immunoprecipitation (IP), a polyclonal anti-follistatin antibody is applied. An evaluation of suitable antibodies for IP and immunoblotting is also presented. Furthermore, practically all currently available follistatin standards were investigated. The detection limit of the method for black market FS344 in urine is ca 0.1 ng/mL for 10 mL. For a sample volume of 100 μL, an LOD of 5 ng/mL could be achieved for serum. Due to the presence of His-tags an unambiguous differentiation from endogenous follistatin is possible.
Keywords: SDS-PAGE; black market; doping control; follistatin 344; myostatin inhibitor.
© 2019 John Wiley & Sons, Ltd.
PubMed Disclaimer
References
REFERENCES
Phillips DJ, de Kretser DM. Follistatin: a multifunctional regulatory protein. Front Neuroendocrinol. 1998;19(4):287-322.
Schneyer AL, Wang Q, Sidis Y, Sluss PM. Differential distribution of follistatin isoforms: application of a new FS315-specific immunoassay. J Clin Endocrinol Metab. 2004;89(10):5067-5075.
De Kretser DM, Hedger MP, Burger HG. Gonadal peptides: inhibins, activins, follistatin, and Müllerian-inhibiting substance (antimüllerian hormone). In: Jameson JL, De Groot L, eds. Endocrinology: Adult and Pediatric. 7th ed. Philadelphia, PA: Saunders; 2016:2037-2050.
Michel U, Albiston A, Findlay JK. Rat follistatin: gonadal and extragonadal expression and evidence for alternative splicing. Biochem Biophys Res Commun. 1990;173(1):401-407.
Al-Zaidy SA, Sahenk Z, Rodino-Klapac LR, Kaspar B, Mendell JR. Follistatin gene therapy improves ambulation in Becker muscular dystrophy. J Neuromuscul Dis. 2015;2(3):185-192.
LinkOut - more resources
Full Text Sources
Wiley