Universität zu Köln

Vogel, Matthias, Blobel, Mike, Thomas, Andreas ORCID: 0000-0003-1199-0743, Walpurgis, Katja, Schaenzer, Wilhelm, Reichel, Christian and Thevis, Mario (2014). Isolation, Enrichment, and Analysis of Erythropoietins in Anti-Doping Analysis by Receptor-Coated Magnetic Beads and Liquid Chromatography Mass Spectrometry. Anal. Chem., 86 (24). S. 12014 - 12022. WASHINGTON: AMER CHEMICAL SOC. ISSN 1520-6882

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Abstract

Human erythropoietin (hEPO) is an erythropoiesis stimulating hormone frequently employed in antianemia therapy. Its capability to increase the amount of red blood cells however makes hEPO and its derivatives also attractive to dishonest athletes aiming at an artificial and illicit enhancement of their endurance performance. A major objective of the international antidoping fight is the elimination of drug misuse and prevention of severe adverse effects caused by nontherapeutic administrations of highly potent drugs. The emergence of novel and innovative erythropoietin-mimetic agents (EMAs) has been continuously growing in the last years, and the option of using dedicated monoclonal antibodies (mAb) for analytical and sample preparation approaches is gradually reaching limits. In the present study the common ability and property of all EMAs, to bind on the human erythropoietin receptor (hEPOR), is therefore exploited. An alternative methodology to isolate and analyze EMAs, in particular endogenous EPO and the recombinant forms EPOzeta, darbepoetin alfa, and C.E.R.A., from human urine is described, employing conventional ultrafiltration for preconcentration of the target analytes followed by EMA-specific isolation via hEPOR-bound magnetic beads. Analytical data were generated by means of gel-based electrophoretic analysis and nanoliquid chromatography/high resolution/high accuracy tandem mass spectrometry. Limits of detection enabled by the established sample preparation protocols were approximately 20 pg/mL for EPOzeta, 30 pg/mL for darbepoetin alfa, and 80 pg/mL for C.E.R.A.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Vogel, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Blobel, Mike UNSPECIFIED UNSPECIFIED UNSPECIFIED Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Walpurgis, Katja UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaenzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED Reichel, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-420140
DOI:	10.1021/ac5024765
Journal or Publication Title:	Anal. Chem.
Volume:	86
Number:	24
Page Range:	S. 12014 - 12022
Date:	2014
Publisher:	AMER CHEMICAL SOC
Place of Publication:	WASHINGTON
ISSN:	1520-6882
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language RECOMBINANT-HUMAN-ERYTHROPOIETIN; CHRONIC KIDNEY-DISEASE; STIMULATING PROTEIN NESP; DARBEPOETIN-ALPHA; AFFINITY CHROMATOGRAPHY; EPOETIN-ALPHA; ANEMIA; PHARMACOKINETICS; PHYSIOLOGY; BINDING Multiple languages Chemistry, Analytical Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/42014