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

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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).

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.

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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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Authors and Affiliations

  1. Department of Chemistry-Biomedical Center, Uppsala University, Box 599, 751 24, Uppsala, Sweden

    Maria Lönnberg & Kristina Råsbo

  2. Department of Chemistry, Environment and Feed Hygiene, The National Veterinary Institute (SVA), Uppsala, and Division of Analytical Pharmaceutical Chemistry, Biomedical Center, Uppsala University, 751 23, Uppsala, Sweden

    Ulf Bondesson

  3. L.C.H., Laboratoire des Courses Hippiques, 15 rue de Paradis, 91370, Verrières le Buisson, France

    Florence Cormant, Patrice Garcia, Yves Bonnaire, Marie-Agnes Popot & Ludovic Bailly-Chouriberry

  4. MAIIA Diagnostics, 751 38, Uppsala, Sweden

    Jan Carlsson & Niclas Rollborn

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  1. Maria Lönnberg
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  2. Ulf Bondesson
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  3. Florence Cormant
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  4. Patrice Garcia
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  5. Yves Bonnaire
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  6. Jan Carlsson
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  7. Marie-Agnes Popot
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  9. Kristina Råsbo
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  10. Ludovic Bailly-Chouriberry
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Correspondence to Maria Lönnberg.

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Lönnberg, M., Bondesson, U., Cormant, F. et al. Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test. Anal Bioanal Chem 403, 1619–1628 (2012). https://doi.org/10.1007/s00216-012-5972-0

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  • DOI: https://doi.org/10.1007/s00216-012-5972-0

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