Effect of exercise induced proteinuria and hypoxia on the analysis of rEPO

Code: 242B05SV

ERA analysis has been repeatedly legally challenged. One of the criticised points is potential protein overload related to exercise induced proteinuria. Although exercise induced proteinuria has been investigated for IEF and SDS-PAGE and samples are immunopurified, no standardized data are available for the currently used SAR-PAGE method.

Although both methods apply the same measurement principle, which is basically separation by migration behaviour based on molecular weight and posttranslational modifications such as glycosylation or pegylation of the molecule, the detergent is different. In addition, the analysis method evolved with the introduction of the biotinylated version of the AE7A5 Anti-EPO antibody being now used for the analysis, and no data exist which can exclude the possibility that contents of exercise urines might cross-react with, for example, the Streptavidin-HRP complex which is used for the visualization by chemiluminescence.

Further, it has not been demonstrated whether high intensity exercise in a hypoxic environment could further affect the ERA analysis, for example by a higher excretion of erythropoietin. Therefore, it is planned to investigate urine samples from 30 athletes who are subjected to an exercise protocol on a treadmill or cycling ergometer known to induce proteinuria. In addition, urine samples from 10 athletes following an acute hypoxic exercise protocol will be collected and examined.