Capillary Electrophoresis in Erythropoietin Quality Control

Abstract

Introduction: Recombinant human erythropoietin is a glycoprotein hormone that stimulates erythroid cell proliferation, produced at the Center for Molecular Immunology from Chinese hamster ovary cells. The production process is controlled by analytical techniques that evaluate, among other quality attributes, identity and potency as described in the European Pharmacopoeia. On the other hand, the refinement, reduction and replacement (3R) of bioassays by less variable and more accurate physicochemical methods is a common objective of the scientific community, regulatory authorities and pharmaceutical companies.

Objective: To evaluate the capillary electrophoresis technique described for the identification of erythropoietin, as a method for the determination of its bioactivity using the I-number model.

Methods: We performed the assembly of the I-number method as an alternative method to the in vivo assay and studied precision parameters and potency accuracy of the international physicochemical standard for erythropoietin (CRS 1.0). Potency was evaluated in ten batches of active pharmaceutical ingredient produced by this method and accuracy was determined with respect to the conventional method.

Results: The type profile of the international standard was obtained, with an I-number value similar to that reported in the literature. Variability of less than 1 % and accuracy of 99 % of potency of the standard was found by the I-number method. The potency specification was met in all evaluated batches and an accuracy between 107 % and 120 % was found.

Conclusions: It was demonstrated that the I-number method constitutes a suitable candidate for the replacement of the in vivo assay in the determination of erythropoietin potency which, in addition to considerably reducing costs, reduces variability and increases measurement accuracy; which has a direct impact on the safety and efficacy of the product.