Universität zu Köln

Arsene, Cristian, Schulze, Dirk, Roethke, Anita ORCID: 0000-0002-3963-5287, Thevis, Mario and Henrion, Andre (2018). Growth hormone isoform-differential massspectrometry for doping control purposes. Drug Test. Anal., 10 (6). S. 938 - 947. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

Mass spectrometry (MS) allows for monitoring growth hormone (GH) isoform compositions at high specificity. It is demonstrated that this capability can be used to reliably detect alterations as elicited by (putative) doping with 22kDa-GH. Sample treatment consists of enzymatic protein cleavage, followed by 2-step liquid chromatography clean-up, prior to analysis by MS. The protocol does not depend on antibodies for analyte extraction at any stage. Therefore, MS opens an opportunity for independent confirmation, if combined with the antibody-based isoform differential test presently used in practice. To check the fitness-for-purpose of this concept, GH-free serum was spiked with pure 22kDa- and 20kDa-GH covering a representative range of concentrations (0.5-9.4g/L), while the 22kDa fraction was within a range of 80%-85%, or at 100%, the latter simulating an administration of 22kDa-GH. Mean deviation of 22kDa-fractions found was within less than 3% for samples with total GH>= 1 g/L . Beyond this, results by antibody-free isoform-differential MS, as described, were in line with those of the World Anti-Doping Agency-approved antibody-based test for 18 native sera and 3 positive controls. In this context, relating 22kDa-GH to total-GH rather than 22kDa+20kDa was considered as an alternative strategy to earlier approaches. However, 20kDa-GH as an additional measurand, next to 22kDa- and total-GH, provides useful extra information, as it directly indicates the presence or absence of a non-22kDa-GH form.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Arsene, Cristian UNSPECIFIED UNSPECIFIED UNSPECIFIED Schulze, Dirk UNSPECIFIED UNSPECIFIED UNSPECIFIED Roethke, Anita UNSPECIFIED orcid.org/0000-0002-3963-5287 UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED Henrion, Andre UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-184726
DOI:	10.1002/dta.2350
Journal or Publication Title:	Drug Test. Anal.
Volume:	10
Number:	6
Page Range:	S. 938 - 947
Date:	2018
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1942-7611
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MASS-SPECTROMETRY; SPORTS; QUANTIFICATION; IMMUNOASSAYS; ABUSE; SERUM; KDA; GH Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18472