Abstract
While firstand second-generation recombinant erythropoietins (EPOs) are detectable in urine, we showed that analysis in blood is more appropriate for the third-generation, continuous erythropoietin receptor activator (CERA), due to its poor excretion in urine. However, isoelectric focusing (IEF) of EPO in serum or plasma is not as simple as in urine since it requires an initial preparative step by immunoaffinity chromatography. That is why enzyme-linked immunosorbent assay (ELISA) and IEF are used as screening and confirmation tests for CERA detection, respectively. Due to the observation of false negative results of ELISA, we have developed a fast method for preparing plasma or serum samples, making IEF as easy to use in blood as in urine. While preparation by immunoaffinity is kept for confirmation analysis, this convenient method is proposed for screening analysis. In addition to CERA, Darbepoetin alfa is very easily identified in blood due to its typical IEF profile. Identification of first-generation EPO drugs will require criteria specific for blood. In comparison with urine, EPO analyses in blood appear beneficial in terms of sensitivity, stability during physical exercise, and prevention of sample adulteration at collection. Anti-doping control of erythropoietin (EPO) is based on the differentiation of natural endogenous from recombinant hormones by their isoelectric profiles.[1] SDS (sodium dodecyl sulfate) electrophoresis is a useful complementary method.[2] Both methods were developed for urine samples. While firstand second-generation recombinant EPOs are detectable in urine, we showed that analysis in blood is more appropriate for the thirdgeneration, continuous erythropoietin receptor activator (CERA), due to its poor excretion in urine.[3] However, isoelectric focusing (IEF) of EPO in serum or plasma is not as simple as in urine since it requires an initial preparative step by immunoaffinity chromatography.[4] We thus developed a convenient method (more than 20 samples can be treated in parallel) for preparing plasma or serum samples, making IEF as easy to use in blood as in urine.
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