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

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS Journal of Chromatography B Pub Date : 2025-02-10 DOI:10.1016/j.jchromb.2025.124518

Nupur Garg, Sanjay Mendiratta, Gurminder Bindra, Sakshi Gehlot, Krutika Goswami, Charu Mehra Kamal

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Abstract

Darbepoetin alfa is a new generation recombinant human erythropoiesis stimulating agent having extended half-life as compared to recombinant human erythropoietin. It is given as a life-saving drug for anemic disorders caused by chronic kidney diseases, cancer, etc. With the presence of commercial formulations of darbepoetin biosimilars in the market and non-availability of any established method in literature or testing guidelines in any compendia for their purity and quality, this research aims to develop a sensitive and robust method to determine the aggregate impurities by using size exclusion chromatographic technique. The method was optimized for sharp and high-resolution monomer peak of darbepoetin and other interferences which was further validated according to ICH Q2(R2) guidelines. The developed method was further tested on the commercial samples to verify the same. Additionally, stress testing of samples was done to simulate the after-production exposure and stability assessment. The results demonstrated that the method for analysing aggregates in darbepoetin is complying the criteria of being sensitive, specific, accurate, robust and reliable.

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通过开发单一有效的分析方法估算 Darbepoetin alfa 制剂中的蛋白质聚集物

达贝泊汀是新一代重组人促红细胞生成素，与重组人促红细胞生成素相比，其半衰期延长。它是一种救命药，用于治疗由慢性肾脏疾病、癌症等引起的贫血症。由于市场上存在达贝泊丁生物类似药的商业配方，并且没有任何文献或任何药典中的测试指南可用于其纯度和质量，本研究旨在开发一种灵敏且可靠的方法，通过粒径排除色谱技术来确定聚集杂质。根据ICH Q2（R2）指南对该方法进行了优化，以获得达贝泊汀的高分辨率单体峰和其他干扰。对所开发的方法进行了进一步的商业样品测试，以验证该方法的有效性。此外，还对样品进行了应力测试，模拟生产后的暴露和稳定性评估。结果表明，该方法具有灵敏、特异、准确、稳健、可靠的特点。

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.