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Multivariate statistics for the differentiation of erythropoietin preparations based on intact glycoforms determined by CE-MS

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Abstract

Owing to the increasing number of erythropoietin biosimilars being approved, the comparison of different erythropoietin preparations in the pharmaceutical area is gaining in importance. Erythropoietin has a distinct natural heterogeneity arising from its glycosylation, which shows strong composition variations. This heterogeneity increases the complexity of the analysis of erythropoietin considerably, but may also be used to distinguish different preparations. Here, a method is presented for the differentiation of various erythropoietin preparations by capillary electrophoresis–mass spectrometry and the subsequent application of multivariate statistics. Relative peak areas of selected intact erythropoietin isoforms were used as variables in principal component analysis and hierarchical agglomerative clustering. Both of these strategies were suited for the clear differentiation of all erythropoietin preparations, including marketed products and preproduction preparations, which differ in the manufacturer, the production cell line, and the batch number. By this means, even closely related preparations were distinguished on the basis of the combined information on the antennarity, the sialoform, and the acetylation of the observed isoforms.

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Acknowledgments

The authors acknowledge financial support in the course of the announcement “Sicherung der Warenketten” of the Federal Ministry of Education and Research (BMBF) within the scope of the program “Forschung für die zivile Sicherheit” of the Federal Government.

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Authors and Affiliations

  1. Chemistry Department, Aalen University, Beethovenstrasse 1, 73430, Aalen, Germany

    Angelina Taichrib, Markus Pioch & Christian Neusüß

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  1. Angelina Taichrib
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  2. Markus Pioch
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  3. Christian Neusüß
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Correspondence to Christian Neusüß.

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Taichrib, A., Pioch, M. & Neusüß, C. Multivariate statistics for the differentiation of erythropoietin preparations based on intact glycoforms determined by CE-MS. Anal Bioanal Chem 403, 797–805 (2012). https://doi.org/10.1007/s00216-012-5924-8

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  • DOI: https://doi.org/10.1007/s00216-012-5924-8

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