Formulation, bioprocessing, and manufacturing of viral vectors in unique dosage forms

Though the market for biological drugs has rapidly expanded in recent years, most formulations still require the cold chain for sufficient thermostability, making these therapies costly. When looking to improve the overall shelf-life stability profile of biological therapeutics, solid-state stabilization techniques are commonly used. These techniques have long been dominated by dry preparations like lyophilization and spray drying, where low residual moisture (RM) content is necessary for stability. However, these techniques are generally expensive, requiring long development and processing times, and are typically inaccessible to low-income countries. Our lab has developed a film technology that can stabilize biologics at ambient temperatures, with relatively high RM, especially when compared to existing lyophilization and spray dried preparations. Here, we investigate the importance of the film drying parameters and storage relative humidity (RH) on the resulting film’s RM, and how this impacts the thermostability profile of the film. When evaluating Adenovirus (Ad) and Adeno-Associated Virus Serotype 9 (AAV9), we found that a controlled equilibrium drying at high RH (52.5% RH) was optimal for long-term thermostability. We also found that Ad was sensitive to both drying and storage RH, while AAV9 was less sensitive in comparison. After investigating the film matrix stability, we determined that the amorphous film matrix itself was relatively stable, and that virus degradation in the film matrix could be due to aggregation and chemical degradation of virus within the microenvironment of the film. Lastly, another method to improve to biological therapies is PEGylation. This technique aims to improve the stability of a product in vivo, as it typically increases the half-life of biologics in the body. However, to properly characterize a successful PEGylation, assays are needed to monitor and verify the conjugation to ensure consistency and repeatability. Here, we have outlined a small-scale PEGylation process that can be easily implemented at the bench scale, which allows researchers to quickly conjugate and evaluate their given biologic for efficacy. To this end, the goal is to provide a blueprint for efficiently developing a PEGylated biologic to be evaluated in preclinical studies. Furthermore, we have outlined the minimum protein or vector requirements needed to characterize the PEGylation process, in the event that material restraints are an issue.