The Kennedy College of Science, Department of Chemistry, is hosting a Ph.D. research proposal defense by Mahsa Aghaei on August 25, 2025. The focus of the research is on understanding the importance of disulfide bonds in maintaining the stability, activity, and safety of therapeutic proteins. Disulfide bonds play a crucial role in protein folding and function, but their dynamic nature can lead to structural changes that affect the quality of biologic drugs. This study aims to enhance the design, manufacturing, and storage of complex biotherapeutics by investigating the dynamics of disulfide bonds in therapeutic proteins.
The first aim of the study is to evaluate the disulfide bond dynamics in a fusion protein consisting of a knob-into-hole (KiH) Fc and the IdeS protease. Different configurations of the IdeS-Fc fusion protein will be analyzed to understand how they impact disulfide bond architecture and cysteine status. The study will involve storing the constructs at different temperatures and time points to observe potential disulfide bond shuffling. By using a validated LC-MS-based platform, the research aims to monitor disulfide bond rearrangements and free cysteine modifications with high sensitivity. The findings from this aim will provide valuable insights into the stability of the fusion protein, guiding the design of more stable and effective therapeutic proteins.
The second aim of the study expands the investigation to assess how manufacturing variables, such as host cell systems and bioprocessing parameters, influence the disulfide bond configuration of a scFv-TCR fusion protein. By comparing proteins produced in different cell culture environments, the research will shed light on how upstream process conditions affect the redox state, disulfide bond formation, and overall structural integrity of the protein. This knowledge is crucial for optimizing production workflows to ensure consistent quality and functional stability of biotherapeutics, reducing variability and potential immunogenicity.
Overall, this research aims to uncover the mechanisms governing disulfide bond stability and dynamics in complex therapeutic proteins. The findings will have significant implications for biopharmaceutical development, providing valuable insights for designing more stable constructs, refining quality control techniques, and establishing bioprocessing standards to minimize modifications that can compromise drug efficacy and safety. Ultimately, this study will contribute to the safe and effective application of biologic therapeutics, driving innovation in protein engineering and supporting regulatory science in biomanufacturing. Interested students and faculty members are encouraged to attend the research proposal defense to learn more about this important research.
Takeaways:
– Investigating the dynamics of disulfide bonds in therapeutic proteins is crucial for enhancing the design and manufacturing of biotherapeutics.
– Understanding how manufacturing variables influence disulfide bond configuration can help optimize production workflows and ensure consistent quality of biologic drugs.
– This research aims to advance the application of biologic therapeutics by providing insights into improving stability, refining quality control, and minimizing modifications that compromise drug efficacy and safety.
Tags: cell culture, regulatory, protein stability, upstream, biomanufacturing, quality control, protein engineering
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