Erythropoietin (EPO) immunoaffinity columns--a powerful tool for purifying EPO and its recombinant analogues.
The sample preparation method preceding the urinary erythropoietin (EPO) doping test is based on several concentration and ultrafiltration steps. In order to improve the quality of isoelectric focusing (IEF) gel results and therefore, the sensitivity of the EPO test, new sample preparation methods relying on affinity purification were recently proposed. This article focuses on the evaluation and validation of disposable immunoaffinity columns targeting both endogenous and recombinant EPO molecules in two World Anti-Doping Agency (WADA) accredited anti-doping laboratories. The use of the columns improved the resolution of the IEF profiles considerably when compared with the classical ultrafiltration method, and the columns' ability to ensure the isoform integrity of the endogenous and exogenous EPO molecules was confirmed. Immunoaffinity columns constitute therefore a potent and reliable tool for the preparation of urine samples and their use will significantly improve the sensitivity and specificity of the actual urinary EPO test.
Dehnes
Yvette
Y
Norwegian Doping Control Laboratory, Oslo University Hospital, Trondheimsveien 235, Oslo, Norway. yvette.dehnes@h-lab.no
Lamon
Séverine
S
Lönnberg
Maria
M
D002846:Chromatography, Affinity / Q000379:methods*; D004300:Doping in Sports; D004921:Erythropoietin / Q000302:isolation & purification* / Q000652:urine; D006801:Humans; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods*; D011994:Recombinant Proteins; D015203:Reproducibility of Results
D011994:Recombinant Proteins; D004921:Erythropoietin