Algorithm-Based Liquid Formulation Development Including a DoE Concept Predicts Long-Term Viral Vector Stability

Area of Science:

• Biotechnology
• Biopharmaceutical Formulation
• Viral Vector Stability

Background:

• Amino acid-based formulations show promise in preventing stress-induced degradation of complex molecules, including monoclonal antibodies and viral vectors.
• Adenovirus 5 (Ad5) is a model system for studying viral vector stability under various storage conditions.

Purpose of the Study:

• To evaluate the efficacy of tailored amino acid formulations in protecting viral vectors, specifically Ad5, against thermal stress and during long-term liquid storage.
• To develop a predictive model for viral vector stability using an algorithm-based approach and design of experiments.

Main Methods:

• Utilized an excipient database for algorithm-based amino acid preselection.
• Employed design of experiments (DoE) and a 37°C thermal challenge model to predict long-term storage stability of Ad5.
• Analyzed Ad5 infectivity statistically to identify key amino acids influencing stability.
• Tested Ad5 formulations with selected amino acids for infectivity after 24 months at 5°C.

Main Results:

• Identified specific amino acids significantly enhancing Ad5 stability after 2-3 weeks of liquid storage at 37°C.
• Ad5 formulations with selected amino acids maintained infectivity after 24 months at 5°C.
• Standard viral vector formulations showed a 2-log reduction in infectivity after 3 months and complete loss after 18 months at 5°C.
• Developed an optimized formulation that excluded MgCl2, which was previously considered essential for Ad5.

Conclusions:

• Algorithm-based formulation development is effective for enhancing viral vector stability.
• Tailored amino acid formulations provide superior protection for viral vectors during long-term liquid storage compared to standard methods.
• The developed formulation strategy offers a robust and efficient approach for biopharmaceutical development.