Rapid detection of erythropoiesis-stimulating agents in urine and serum

• Published in: Analytical biochemistry Vol. 420; no. 2; pp. 101 - 114
• Main Authors: Lönnberg, Maria, Andrén, Maria, Birgegård, Gunnar, Drevin, Malin, Garle, Mats, Carlsson, Jan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-01-2012
• Subjects: Adult; Affinity Chromatography; agglutinins; antibodies; Antibodies - immunology; Blood Chemical Analysis - instrumentation; Blood Chemical Analysis - methods; blood serum; chromatography; Doping Control; EPO; EPOerythropoietin; Erythropoiesis - drug effects; Erythropoietin; Erythropoietin - blood; Erythropoietin - immunology; Erythropoietin - metabolism; Erythropoietin - urine; Female; glycosylation; hamsters; Hematinics - blood; Hematinics - pharmacology; Hematinics - urine; human cell lines; Humans; Image Scanner; Immunoassay; immunoassays; Lateral flow immunoassay; Male; Medicin och hälsovetenskap; Micro-Column; Microcolumn; Middle Aged; Polyethylene Glycols; Protein Isoforms; Protein Isoforms - blood; Protein Isoforms - immunology; Protein Isoforms - metabolism; Protein Isoforms - urine; rapid methods; soot; therapeutics; Time Factors; Urinalysis - instrumentation; Urinalysis - methods; urine; WGA; wheat germ; Wheat Germ Agglutinins - metabolism; Young Adult; Protein isoforms; WGA; Microcolumn; Erythropoietin; EPO; Affinity chromatography; Lateral flow immunoassay; Doping control; Image scanner
• ISSN: 0003-2697; 1096-0309; 1096-0309
• DOI: 10.1016/j.ab.2011.09.021

A rapid and easy-to-use test kit, EPO WGA MAIIA, which can be used for distinguishing various endogenous human erythropoietins (hEPOs) and several recombinant hEPO 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 using anti-EPO carbon black nanostrings and image scanning for quantification. All of the reactions take place along the porous layer of a lateral flow microcolumn containing WGA and anti-EPO zones. The presence of molecules resembling hEPOs, such as Mircera, was detected by the aberrant affinity interaction with the antibody zone on the strip. It was possible to distinguish nine recombinant hEPOs expressed in hamster and human cell lines, as well as Aranesp and Mircera, from endogenous urine hEPO. The required amount of EPO in the samples, a few picograms, is very low compared with 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.