[Determination methods for erythropoietin receptor activator in human urine].
In the present study, isoelectronic focusing with different pH gradients (pH 3-5, 2-6) or migrating distances (8.5, 12 and 17 cm) and SDS-PAGE was used to separate continuous erythropoietin receptor activator (CERA), recombinant human erythropoietin (rhEPO), darbepoetin and endogenous EPO spiked in human urine with 37 degrees C overnight incubation. Double blotting and chemiluminescent visualization were used to detect the IEF and SDS-PAGE profiles. The bands of CERA profile were detected and well separated from the endogenous EPO and the other two EPO preparations with both SDS-PAGE and the IEF method using a gradient pH 3-5 and a migrating distance of 17 cm, and a significant particular band of CERA profile was found in the IEF result. These preliminary results indicated that the methods were reliable and reproducible for detecting CERA, and could be used as a routine procedure for anti-doping analysis.
Xing
Yan-Yi
YY
National Anti-doping Agency, Beijing 100029, China. xingyanyi@tom.com
Zhang
Li-Si
LS
Xu
You-Xuan
YX
Wu
Mou-Tian
MT
Wang
Shan
S
D004591:Electrophoresis, Polyacrylamide Gel; D004921:Erythropoietin / Q000652:urine*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D011092:Polyethylene Glycols; D011994:Recombinant Proteins
D011994:Recombinant Proteins; C508420:continuous erythropoietin receptor activator; D004921:Erythropoietin; D011092:Polyethylene Glycols