Reichel, Christian and Thevis, Mario (2012). Detection of EPO-Fc fusion protein in human blood: Screening and confirmation protocols for sports drug testing. Drug Test. Anal., 4 (11). S. 818 - 830. HOBOKEN: WILEY-BLACKWELL. ISSN 1942-7603
Full text not available from this repository.Abstract
The neonatal Fc receptor (FcRn) has been under investigation for several years as a pharmaceutical drug target. Clinical studies have shown that fusion proteins consisting of human recombinant erythropoietin (rhEPO) and the Fc-part of IgG can be transported after pulmonary administration via FcRn across the airway epithelium to the blood stream. So far, no clinically approved pharmaceutical formulation of EPO-Fc is available. Since various forms of recombinant erythropoietins have been frequently misused by athletes as performance-enhancing agents, EPO-Fc might play a similar role in sports in the future. In order to investigate the detectability of EPO-Fc in human blood, different strategies were tested and developed. Only two of them fulfilled the necessary requirements regarding sensitivity and specificity. A rapid protocol useful for screening purposes first enriches EPO-Fc from human serum via high capacity protein A beads and subsequently detects EPO-Fc in the eluate with a commercial EPO ELISA kit. The limit of detection (LOD) of the method is about 5 pg (45 amol) EPO-Fc and is independent of the serum volume used. For screening and/or confirmation purposes a second protocol was evaluated, which consists of a fast EPO immunopurification step followed by sodium dodecyl sulfate or sarcosyl polyacrylamide gel electrophoresis (SDS-PAGE, SAR-PAGE) and Western double-blotting with chemiluminescence detection a method already established in routine EPO anti-doping control. The latter strategy allows the detection of EPO-Fc in serum together with all other recombinant erythropoietins and with an identical LOD (5 pg/45 amol) as for the rapid screening protocol. Copyright (C) 2012 John Wiley & Sons, Ltd.
Item Type: | Journal Article | ||||||||||||
Creators: |
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URN: | urn:nbn:de:hbz:38-479494 | ||||||||||||
DOI: | 10.1002/dta.1381 | ||||||||||||
Journal or Publication Title: | Drug Test. Anal. | ||||||||||||
Volume: | 4 | ||||||||||||
Number: | 11 | ||||||||||||
Page Range: | S. 818 - 830 | ||||||||||||
Date: | 2012 | ||||||||||||
Publisher: | WILEY-BLACKWELL | ||||||||||||
Place of Publication: | HOBOKEN | ||||||||||||
ISSN: | 1942-7603 | ||||||||||||
Language: | English | ||||||||||||
Faculty: | Unspecified | ||||||||||||
Divisions: | Unspecified | ||||||||||||
Subjects: | no entry | ||||||||||||
Uncontrolled Keywords: |
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URI: | http://kups.ub.uni-koeln.de/id/eprint/47949 |
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