Estimation of protein aggregates in Darbepoetin alfa formulations by developing a single validated analytical method

Area of Science:

• Biopharmaceutical analysis
• Chromatographic techniques
• Drug quality assessment

Background:

• Darbepoetin alfa, a long-acting erythropoiesis-stimulating agent, treats anemia in chronic kidney disease and cancer.
• Commercial darbepoetin alfa biosimilars lack established purity and quality testing methods.
• Ensuring the quality of biosimilars is critical for patient safety and therapeutic efficacy.

Purpose of the Study:

• To develop and validate a sensitive and robust size exclusion chromatography (SEC) method for determining aggregate impurities in darbepoetin alfa biosimilars.
• To establish a reliable analytical approach for quality control of available darbepoetin alfa biosimilar formulations.

Main Methods:

• Size exclusion chromatography (SEC) was employed to separate and quantify aggregate impurities.
• Method optimization focused on achieving high resolution for the darbepoetin monomer peak and minimizing interferences.
• Validation was performed according to International Council for Harmonisation (ICH) Q2(R2) guidelines.

Main Results:

• The developed SEC method demonstrated high sensitivity, specificity, accuracy, robustness, and reliability.
• The method successfully analyzed aggregate impurities in commercial darbepoetin alfa biosimilar samples.
• Stress testing confirmed the method's suitability for stability assessment and post-production exposure simulation.

Conclusions:

• A validated SEC method provides a reliable tool for assessing aggregate impurities in darbepoetin alfa biosimilars.
• This method supports the quality control and assurance of darbepoetin alfa biosimilar products.
• The findings contribute to ensuring the safety and efficacy of anemia treatments utilizing darbepoetin alfa biosimilars.