Abstract

Human growth hormone (GH) is prohibited in sports by the World Anti-Doping Agency (WADA) and two different approaches for a doping control method are currently discussed. The Strasburger test is presently used as doping control method and the marker test measuring the indirect markers IGF-I and Procollagen III by Sönksen et al. is discussed as complementation. The assay that is presented here could be used as confirmation assay in combination with the Strasburger test. The method based on 2D-PAGE was validated and a reference population was analyzed to establish a discrimination limit. As a proof of principle, patient samples were prepared and were clearly positive 2.5 h after s.c. injection. After 12-16 h, three out of the five samples still had values higher than the discrimination limit. Four variants are detected in endogenous samples (22 kDa GH, splice variant, phophorylated GH, and glycosylated GH) and the mass spectrometric identification of the glycosylated variant is presented here. For comparison and to test the combination of the Strasburger test and the 2D-PAGE method the patient samples were analyzed with both tests. The results showed that a combination of both tests can improve the overall performance of growth hormone analysis in doping control. Furthermore an antibody alternative was tested and the sensitivity of the 2D-PAGE method was shown to be improved by the use of a secondary AMDEXTM antibody. Verf.-Referat