A Reference Standard for Analytical Testing of Erythropoietin

Huiping Tu,Ben Cowper,Mark Hasselberg,Rebecca Potts,Mark Verdecia,Chris Burns,Kevin Carrick,Fouad Atouf

doi:10.1007/s11095-022-03213-1

Abstract

PurposeErythropoietin (EPO) is a 165 amino acid protein that promotes the proliferation of erythrocytic progenitors. A decrease in endogenous EPO production causes anemia that can be treated with recombinant Human EPO (rHuEPO).ObjectiveTo ensure the safety and efficacy of the rHuEPO, manufacturers must use analytical methods to demonstrate similarity across batches and between different products. To do this they need reference standards to validate their equipment and methods.MethodWe used peptide mapping, size-exclusion chromatography, glycoprofiling, and isoelectric focusing to analyze a rHuEPO reference standard.ResultsCharacterization demonstrates that our rHuEPO reference standard meets the criteria for quality.ConclusionThe rHuEPO reference standard is fit for purpose as a tool for validating system suitability and methods.

Highlights

Manufacturers of erythropoietin (EPO) biosimilars must demonstrate that their product is highly similar to and has no clinically meaningful differences from that of an already licensed reference product (RP). [1] This requirement assumes that the biosimilar manufacturer has access to batches of RP, as well as an in-depth understanding of the relevant critical quality attributes (CQAs)
Peptide mapping was performed on the reformulated United States Pharmacopeial Convention (USP) recombinant human erythropoietin (rHuEPO)-reference standard (RS) to demonstrate its similarity to the source bulk drug substance (DS)
Recombinant human erythropoietin is prescribed to treat renal anemia associated with chronic renal failure and chemotherapy treatments

Summary

Introduction

Manufacturers of erythropoietin (EPO) biosimilars must demonstrate that their product is highly similar to and has no clinically meaningful differences from that of an already licensed reference product (RP). [1] This requirement assumes that the biosimilar manufacturer has access to batches of RP, as well as an in-depth understanding of the relevant critical quality attributes (CQAs). [3, 4] The biosimilar manufacturer will need to develop its own manufacturing processes (e.g., different cell lines, raw materials, equipment, procedures, process controls, and acceptance criteria). Under these circumstances, it can significantly benefit the biosimilar manufacturer to have access to a public reference standard (RS) for EPO that can be used to validate methods and qualify systems. An RS cannot substitute for an RP for this purpose, but it can be useful for establishing the assays necessary for evaluating the similarity of the candidate biosimilar These assays are highly scrutinized by regulators because they serve as the basis for any claims of biosimilarity. In the United States, comprehensive comparative analytical data are necessary to demonstrate biosimilarity and interchangeability under section 351(k) of the Public Health Service Act, and a “Marketing Authorisation Application” is required under Article 10(4) of Directive 2001/83/EC in Europe. [5, 6] The results of the comparability exercise need to be of such quality as to allow the regulatory agency to draw definitive conclusions about the similarity of the physicochemical, biological, and pharmacological attributes between the therapeutic product under

Methods

Results

Conclusion