Rapid detection of erythropoiesis-stimulating agents in urine and serum
2012 (English)In: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 420, no 2, 101-114 p.Article in journal (Refereed) Published
A rapid and easy-to-use test kit, EPO WGA MAIIA, which can be used for distinguishing various endogenous human erythropoietins (hEPO) and several recombinant hEPOs and EPO analogues, has been evaluated. The test is based on chromatographic separation of the glycosylated isoforms of EPO using wheat germ agglutinin (WGA), and a sensitive immunoassay utilizing anti-EPO carbon black nanostrings and image scanning for quantification. All the reactions take place along the porous layer of a lateral flow micro-column containing WGA and anti-EPO zones. The presence of molecules resembling hEPO, like Mircera, was detected by the aberrant affinity interaction with the antibody zone on the strip. It was possible to distinguish nine recombinant hEPO expressed in hamster and human cell-lines, and also Aranesp and Mircera, from endogenous urine hEPO. The required amount of EPO in the samples, a few pg, is very low compared to other methods for EPO isoform identification. This EPO isoform determination method opens the possibility to monitor recombinant EPO therapy for clinical research and seems to be a valuable candidate to the arsenal of EPO doping control tests.
Place, publisher, year, edition, pages
2012. Vol. 420, no 2, 101-114 p.
EPOerythropoietin, Affinity Chromatography, Doping Control, Image Scanner, Lateral-Flow Immunoassay, Micro-Column, Protein Isoforms, WGA
IdentifiersURN: urn:nbn:se:uu:diva-160163DOI: 10.1016/j.ab.2011.09.021ISI: 000297482700001OAI: oai:DiVA.org:uu-160163DiVA: diva2:450598