Purification of Modified Therapeutic Proteins Available on the Market: An Analysis of Chromatography-Based Strategies

This review article comprehensively analyzes the chromatography-based purification strategies employed for modified therapeutic proteins that have received regulatory approval and are currently commercialized. The paper highlights that protein modification, through covalent attachment to various moieties like PEG, Fc-domains, lipids, albumin, and glycosides, significantly enhances the pharmacokinetic properties, stability, and therapeutic efficacy of proteins. However, these modifications also introduce complexity into the purification process, necessitating robust and selective chromatographic methods. The authors emphasize that chromatography is the gold standard in downstream processing for therapeutic proteins due to its high resolution and robustness. The review delves into specific modification types, including PEGylation, Fc-fusion, and lipidation, detailing the purification process parameters such as chromatographic technique, stationary phase, and elution mode. It also uses coagulation factor IX as a case study to illustrate how modification type profoundly influences chromatographic process development. The article provides a critical analysis of successful purification strategies and offers guidance for selecting appropriate chromatography-based purification processes for novel modified therapeutic proteins, addressing current challenges in developing more efficient methods.

This review article comprehensively analyzes the chromatography-based purification strategies employed for modified therapeutic proteins that have received regulatory approval and are currently commercialized. The paper highlights that protein modification, through covalent attachment to various moieties like PEG, Fc-domains, lipids, albumin, and glycosides, significantly enhances the pharmacokinetic properties, stability, and therapeutic efficacy of proteins. However, these modifications also introduce complexity into the purification process, necessitating robust and selective chromatographic methods. The authors emphasize that chromatography is the gold standard in downstream processing for therapeutic proteins due to its high resolution and robustness. The review delves into specific modification types, including PEGylation, Fc-fusion, and lipidation, detailing the purification process parameters such as chromatographic technique, stationary phase, and elution mode. It also uses coagulation factor IX as a case study to illustrate how modification type profoundly influences chromatographic process development. The article provides a critical analysis of successful purification strategies and offers guidance for selecting appropriate chromatography-based purification processes for novel modified therapeutic proteins, addressing current challenges in developing more efficient methods. Research Background and Significance The purification of therapeutic proteins has evolved into a critical step in biopharmaceutical manufacturing, ensuring product safety, efficacy, and regulatory compliance. Protein modifications such as PEGylation, Fc-fusion, lipidation, albumin conjugation, and glycosylation have been extensively employed to enhance pharmacokinetics, stability, and the