Biology Reference
In-Depth Information
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4
5
6
7
8
9
10
11
+
_
NegQC
Sample Y
Sample X
EPO positive QC
EPO standard
Darbepoetin standard
Figure 1. Electropherogram of rHuEPO and darbepoetin alfa standards; of extracts of urine obtained
from drug-free control volunteers and patients treated with rHuEPO or darbepoetin alfa; and of
extracts of urine from unknown individuals. The anode and cathode sides of the electropherogram are
labeled + and -, respectively. The rHuEPO standard is in lane one and four and the darbepoetin stan-
dard is in lanes five,seven,nine, and 11. The negative quality control sample (lane two) is an extract
of urine from drug-free volunteers showing the normal pattern of endogenous EPO. The EPO posi-
tive quality control sample (lane three) and darbepoetin positive quality control (lane six) are urine
extracts obtained after the administration of rHuEPO or darbepoetin alfa, respectively. Samples X and
Y are urine extracts from two unknown individuals (lanes eight and 10). The bands of endogenous
urinary EPO are faint or absent in lanes six, eight, and 10.
Despite the fact that isoelectric focusing is technically demanding and may
take up to three days to complete, it has generated a great deal of interest
because it is a direct test that can yield conclusive results. Further, the results
of the test and the underlying method have been accepted by CAS [22].
Presently, at least six IOC-accredited laboratories have implemented the test
and several more are working on the method. Investigators are working on
means to improve the test, to expand its use to all 25 IOC-accredited laborato-
ries, and to further define criteria for determining if rHuEPO is present. In
addition, investigators are working on ways to detect erythropoietic proteins
with mass spectrometry.
Darbepoetin alfa detected at Winter Olympic Games of Salt Lake, 2002
As the IOC-accredited laboratory designated to perform the testing at the
Games of Salt Lake, our laboratory began implementing the isoelectric-focus-
ing technique for detecting erythropoietic proteins about eight months before
