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Analytical and Bioanalytical Chemistry

, Volume 403, Issue 6, pp 1619–1628 | Cite as

Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test

  • Maria Lönnberg
  • Ulf Bondesson
  • Florence Cormant
  • Patrice Garcia
  • Yves Bonnaire
  • Jan Carlsson
  • Marie-Agnes Popot
  • Niclas Rollborn
  • Kristina Råsbo
  • Ludovic Bailly-Chouriberry
Original Paper

Abstract

Doping of horses with recombinant human erythropoietin (rHuEPO) to illegally enhance their endurance capacity in horseracing has been reported during the last years. This leads to increased blood viscosity which can result in sudden death and is of concern for the horse welfare. Additionally, the horse can start production of rHuEPO antibodies, which cross-reacts with endogenous equine EPO and can lead to severe anaemia and even death. In this study, a novel micro-chromatographic method, EPO WGA MAIIA, has been tested for the capability in plasma and urine samples to detect administration of erythropoiesis-stimulating agents, like the rHuEPO glycoprotein varieties Eprex and Aranesp, to horses. After administration of 40 IU Eprex kg−1 day−1 to seven horses during 6 days, the presence of Eprex in horse plasma was detected up to 2–5 days after last injection. In urine samples collected from two horses, Eprex was detected up to 3 days. A single injection of Aranesp (0.39 μg/kg) was detected up to 9 days in plasma and up to 8 days, the last day of testing, in the urine sample. The LC-FAIMS-MS/MS system, with 1 day reporting time, confirmed the presence of Eprex up to 1 day after last injection for six out of seven horses and the presence of Aranesp up to 5 days after last injection in plasma samples. The MAIIA system showed to be a promising tool with high sensitivity and extremely short reporting time (1 h).

Fig

The interior of the MAIIA membrane. EPO protein molecules (blue balls) flow through the WGA lectin ligands immobilised in the network structure. Some types of EPO have oligosaccharide structures with higher interaction strength and will be delayed, while other low reacting types will rapidly reach the anti-EPO antibodies immobilised downstream, during the 5 min. reaction time.

Keywords

Aranesp EPO doping control Eprex Equine EPO Micro-chromatography WGA 

Notes

Acknowledgements

The authors thank Maria Andrén, Malin Drevin, Mikael Lönnberg and Trikien Quach for technical assistance, and the Swedish Foundation for Equine Research, Stockholm, Sweden, and MAIIA Diagnostics, Uppsala, Sweden, for support. The authors are indebted to Dr. Jean-Jacques Garin, veterinary surgeon at FNCF, to the horse farm manager in Coye la Forêt and to the staff who participated in drug administration, sampling and horse care.

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Maria Lönnberg
    • 1
  • Ulf Bondesson
    • 2
  • Florence Cormant
    • 3
  • Patrice Garcia
    • 3
  • Yves Bonnaire
    • 3
  • Jan Carlsson
    • 4
  • Marie-Agnes Popot
    • 3
  • Niclas Rollborn
    • 4
  • Kristina Råsbo
    • 1
  • Ludovic Bailly-Chouriberry
    • 3
  1. 1.Department of Chemistry-Biomedical CenterUppsala UniversityUppsalaSweden
  2. 2.Department of Chemistry, Environment and Feed Hygiene, The National Veterinary Institute (SVA), Uppsala, and Division of Analytical Pharmaceutical Chemistry, Biomedical CenterUppsala UniversityUppsalaSweden
  3. 3.L.C.H., Laboratoire des Courses HippiquesVerrières le BuissonFrance
  4. 4.MAIIA DiagnosticsUppsalaSweden

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