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High resolution separation methods for the determination of intact human erythropoiesis stimulating agents. A review

AuthorsGirard, M.; Puerta, Angel de la ; Díez-Masa, José Carlos; Frutos, Mercedes de
Issue Date2012
CitationAnalytica Chimica Acta 713: 7- 22 (2012)
AbstractHuman erythropoietin (hEPO), a hormone involved in the formation of red blood cells, is a 30. kDa glycoprotein with a high carbohydrate content. The production of recombinant hEPO has made possible its widespread therapeutic use and its banned use in competition sports. Methods to analyze EPO and other erythropoiesis stimulating agents (ESAs) are necessary for the characterization and quality control of these biopharmaceuticals and also for doping control. In this paper, high resolution separation methods, namely high performance liquid chromatography (HPLC) and capillary electrophoresis (CE), with special attention to CE-coupled mass spectrometry, are reviewed. The usefulness of these techniques when applied in different modes to separate the glycoprotein isoforms, aggregates or excipients are detailed. In addition, sample preparation methods that have been applied to ESA samples for subsequent determination by HPLC or CE, as well as the potential compatibility of other preparation methods, are discussed. Applications of the HPLC and CE methods regarding regulatory considerations for biopharmaceuticals analysis, with emphasis on biosimilars, and doping control are also included. Finally, limitations of the present methods and their possible solutions are considered. © 2011 Elsevier B.V.
Publisher version (URL)http://dx.doi.org/10.1016/j.aca.2011.11.041
Identifiersdoi: 10.1016/j.aca.2011.11.041
issn: 0003-2670
e-issn: 1873-4324
Appears in Collections:(IQOG) Artículos
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