VALIDATION OF CATION-EXCHANGE CHROMATOGRAPHY METHOD FOR ANALYSIS OF CHARGE HETEROGENEITY OF ANTI-CD20 BIOSIMILAR CANDIDATE ANTIBODY
LESLY PARADINA-FERNÁNDEZ *
Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba.
REYNIER RODRÍGUEZ-HUERTA
Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba.
YAMILET ROMERO-BATISTA
Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba.
LISEL VIÑA-RODRIGUEZ
Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba.
*Author to whom correspondence should be addressed.
Abstract
Appropriate characterization of proteins is vital during the development and production phases of any therapeutic biological product. One of the techniques usually employed for this purpose is the cation-exchange chromatography. Therefore, the adequate validation of this method according to the regulatory agencies becomes essential. In this sense, we report a validation study of a methodology for determine the charges heterogeneity of an anti-CD20 biosimilar candidate antibody (1B8). The assay was validated by examining: robustness, specificity, repeatability, intermediate precision and linearity. The method demonstrated to be robust although, it is necessary a strict control in the mobile phases preparation (pH and molarity). In addition, it was demonstrated the specificity, precision and linearity of this assay. Consequently, it could be considered a suitable tool for the evaluation of 1B8 isoforms profile. Moreover, the present work confirmed that the cation-exchange chromatography is a suitable method in the lot to lot quality control monitoring of final products and active pharmaceutical ingredient of 1B8 monoclonal antibody.
Keywords: Validation, cation-exchange, chromatography, antiCD-20 biosimilar candidate antibody, charge heterogeneity, 1B8