Data from a microdosed recombinant human erythropoietin administration study applying the new biotinylated clone AE7A5 antibody and a further optimized sarcosyl polyacrylamide gel electrophoresis protocol

Data from a microdosed recombinant human erythropoietin administration study applying the new biotinylated clone AE7A5 antibody and a further optimized sarcosyl polyacrylamide gel electrophoresis protocol / Christian Reichel, Damir Erceg, Barbara Lorenc, Veronika Scheiblhofer, Letizia Farmer, Katharina Zanitzer, Thomas Geisendorfer, Günter Gmeiner, Mario Thevis. - (Drug Testing and Analysis (2021) 15 January)

  • PMID: 33450134
  • DOI: 10.1002/dta.2989


Abstract

Erythropoietin (EPO) is a hormone, which stimulates the production of red blood cells. Due to its performance-enhancing effect, it is prohibited by the World Anti-Doping Agency (WADA). In order to reduce the detection window of EPO doping, athletes have been applying low doses of recombinant EPO (e.g., <10 IU/kg body weight, daily or every second day) instead of larger doses twice or more per week (e.g., 30 IU/kg). Microdoses of Retacrit (epoetin zeta), an EPO biosimilar, were administered intravenously and subcutaneously to human males and females. Urine and serum samples were collected and analysed applying the new biotinylated clone AE7A5 EPO antibody and a further optimized sarcosyl polyacrylamide gel electrophoresis (SAR-PAGE) protocol. With the improved protocol, microdosed Retacrit (7.5 IU/kg body weight [BW]) was detectable for at least 52 h after intravenous administration. Detection windows were approximately the same for serum and urine and doubled after subcutaneous administration (~104 h). Previous studies applying different electrophoretic techniques and the not further optimized SAR-PAGE protocol revealed considerably shorter detection windows for recombinant human erythropoietin (rhEPO) microdoses. Because the new biotinylated antibody performed significantly more sensitive than the nonbiotinylated version, the new protocol will improve the sensitivity and hence detectability of recombinant EPO in doping control.

Keywords: SAR-PAGE; doping control; epoetin zeta; erythropoietin; microdosis.

Parameters

Science
Study
Date
21 January 2021
People
Erceg, Damir
Farmer, Letizia
Geisendorfer, Thomas
Gmeiner, Günter
Lorenc, Barbara
Reichel, Christian
Scheiblhofer, Veronika
Thevis, Mario
Zanitzer, Katharina
Country
Austria
Croatia
Germany
Language
English
Other organisations
Children's hospital Srebrnjak
Deutsche Sporthochschule Köln (DSHS) - German Sport University Cologne
European Monitoring Center for Emerging Doping Agents (EuMoCEDA)
Hrvatsko katoličko sveučilište - Catholic University of Croatia
Sv. Katarina Specijalna bolnica - St. Catherine Specialty Hospital
Sveučilište Josipa Jurja Strossmayera u Osijeku (UNIOS) - Josip Juraj Strossmayer University of Osijek
Laboratories
Seibersdorf, Austria: Seibersdorf Labor GmbH Doping Control Laboratory
Analytical aspects
Testing method development
Doping classes
S2. Peptide Hormones, Growth Factors
Substances
Erythropoietin (EPO)
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Abstract
Date generated
22 April 2021
Date of last modification
4 May 2021
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