Universität zu Köln

Thevis, Mario, Thomas, Andreas ORCID: 0000-0003-1199-0743, Geyer, Hans and Schaenzer, Wilhelm (2014). Mass spectrometric characterization of a biotechnologically produced full-length mechano growth factor (MGF) relevant for doping controls. Growth Horm. IGF Res., 24 (6). S. 276 - 281. EDINBURGH: CHURCHILL LIVINGSTONE. ISSN 1532-2238

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Abstract

Objective: Since Goldspink and colleagues identified the expression of the mRNA of an insulin-like growth factor 1 (IGF-1) isoform in response to mechanical stress in 1996, substantial research into the so-called mechano growth factor and its modus operandi followed until today. Promising preclinical results were obtained by using the synthetic, 24-amino acid residues spanning peptide translated from the exons 4-6 of IGF-1Ec (which was later referred to as the mechano growth factor (MGF) peptide), particularly with regard to increased muscle myoblast proliferation. Consequently, the MGF peptide represented a promising drug candidate for the treatment of neuromuscular disorders; however, its misuse potential in sport was also identified shortly thereafter, and the substance (or class of substances) has been considered prohibited according to the regulations of the World Anti-Doping Agency (WADA) since 2005. While various MGF peptide versions have been known to sports drug testing authorities, the occurrence of a 'full-length MGF as offered via illicit channels to athletes or athletes' managers was reported in 2014, arguably being undetectable in doping controls. Methods: An aliquot of the product was obtained and the content characterized by state-of-the-art analytical approaches including gel electrophoretic and mass spectrometric (top-down and bottom-up) sequencing approaches. Upon full characterization, its implementation into modified routine doping controls using ultrafiltration, immunoaffinity-based isolation, and nanoliquid chromatography-high resolution/high accuracy mass spectrometry was established. Results: A protein with a monoisotopic molecular mass of 12264.9 Da and a sequence closely related to IGF-1Ec (lacking the signal- and propeptide moiety) was identified. The C-terminus was found to be modified by the elimination of the terminal lysine and a R109H substitution, With the knowledge of the compound's composition, existing doping control assays targeting peptide hormones such as IGF-1 and related substances were assessed as to their capability to detect the full-length MGF. The analyte was detectable at concentrations of 0.25 ng/mL using adapted routine test methods employing immunoaffinity purification followed by nanoscale liquid chromatography-high resolution/high accuracy (tandem) mass spectrometry. Conclusions: A potentially performance enhancing 'full-length' MGF derivative was identified and successfully implemented into sports drug testing protocols. Future tests are indicated probing for optimized/dedicated detection methods and assessment of efficacy and elimination kinetics of the substance. (c) 2014 Elsevier Ltd. All rights reserved.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Geyer, Hans UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaenzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-421090
DOI:	10.1016/j.ghir.2014.10.004
Journal or Publication Title:	Growth Horm. IGF Res.
Volume:	24
Number:	6
Page Range:	S. 276 - 281
Date:	2014
Publisher:	CHURCHILL LIVINGSTONE
Place of Publication:	EDINBURGH
ISSN:	1532-2238
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language IGF-I; MUSCLE; PEPTIDE; INSULIN; EXPRESSION; STRETCH; CELLS Multiple languages Cell Biology; Endocrinology & Metabolism Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/42109