Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test
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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).
KeywordsAranesp EPO doping control Eprex Equine EPO Micro-chromatography WGA
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.
- 6.Piercy RJ, Swardson CJ, Hinchcliff KW (1998) Erythroid hypoplasia and anemia following administration of recombinant human erythropoietin to two horses. J Am Vet Med Assoc 212(2):244–247Google Scholar
- 12.Macdougall IC, Robson R, Opatrna S, Liogier X, Pannier A, Jordan P, Dougherty FC, Reigner B (2006) Pharmacokinetics and pharmacodynamics of intravenous and subcutaneous continuous erythropoietin receptor activator (C.E.R.A.) in patients with chronic kidney disease. Clin J Am Soc Nephrol 1(6):1211–1215CrossRefGoogle Scholar
- 17.Wide L, Bengtsson C, Berglund B, Ekblom B (1995) Detection in blood and urine of recombinant erythropoietin administered to healthy men. Med Sci Sports Exerc 27(11):1569–1576Google Scholar
- 26.Lasne F, Popot MA, Varlet-Marie E, Martin L, Martin JA, Bonnaire Y, Audran M, de Ceaurriz J (2005) Detection of recombinant epoetin and darbepoetin alpha after subcutaneous administration in the horse. J Anal Toxicol 29(8):835–837Google Scholar
- 30.Bailly-Chouriberry L, Cormant F, Garcia P, Lönnberg M, Szwandt S, Bondesson U, Popot MA, Bonnaire Y (2012) New analytical method based on anti-EPO monolith column for the rHuEPO purification in horse plasma and urine samples. Analyst, DOI: 10.1039/C2AN15662H
- 33.Lönnberg M (2002) Membrane-assisted isoform immunoassay: separation and determination of protein isoforms. Acta Universitatis Upsaliensis Comprehensive Summaries of Uppsala Dissertations, Faculty of Science and Technology, 691 ppGoogle Scholar
- 34.Lonnberg M, Carlsson J (2001) Chromatographic performance of a thin microporous bed of nitrocellulose. J Chromatogr B 763(1–2):107–120Google Scholar