Novel Process Strategies for the Stabilization of Biopharmaceuticals for Parenteral Use

Sharma, Ashutosh (2023) Novel Process Strategies for the Stabilization of Biopharmaceuticals for Parenteral Use. Doctoral thesis, SETU Waterford.

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In the past two decades, biopharmaceuticals have provided a breakthrough in improving the quality of lives of patients with various cancers, autoimmune and genetic disorders etc. With the growing demand of biopharmaceuticals, the need for reducing manufacturing costs is essential without compromising on the safety, quality, and efficacy of products. Freeze-drying is the primary commercial means of manufacturing solid biopharmaceuticals. However, Freeze-drying is an economically unfriendly means of production with long production cycles and heavy capital investment, resulting in high overall costs. This thesis reviews several alternative drying technologies such as continuous Freeze-drying, Spray-drying, Active-freeze-drying, Spray-freeze-drying, PRINT® Technology etc. that have not yet gained popularity for manufacturing parenteral biopharmaceuticals and focuses on assessing the stability of two proteins i.e., Lysozyme by Freeze-drying and Spray-drying, and a commercial therapeutic enzyme by Active-freeze-drying and Spray-drying. The key findings of this thesis showed that lysozyme was a robust protein, and its efficacy was enhanced in the presence of excipients such as sucrose and trehalose post Freeze-drying and Spray-drying. To substantiate experimental results, molecular dynamics simulations were performed that elucidated a conformation change (without unfolding) may have resulted in increased flexibility of the active sites. Furthermore, Enzyme ‘A’, a commercial therapeutic enzyme, was susceptible to process-induced stress post Active-freeze-drying and Spray-drying, thereby, resulting in increased protein aggregation. However, the inclusion of Arg-HCl in the formulation of Enzyme ‘A’ significantly improved the reconstitution time by 63 % and turbidity by 83 % and promoted the suppression of insoluble aggregates post Spray-drying. In agreement with experimental results, molecular dynamics simulations showed that while Arg-HCl was capable of acting as the main stabilizer, it interacted the most with the positively and negatively charged residues on the surface of Enzyme ‘A’ and also acted as a neutral crowder resulting in reduced protein-protein interactions. The last segment of this thesis thoroughly evaluates and discusses the potential of Multi-Angle Dynamic Light Scattering (MADLS) as a 3-in-1 screening tool for the determination of particle size, product concentration and protein aggregation of three proteins including Bovine Serum Albumin (BSA), a commercial monoclonal antibody (mAb) and a therapeutic enzyme. A good calibration curve with an R2 of > 0.95 was obtained between the particle number concentration by MADLS and protein concentration by UV-Vis spectroscopy for the 3 proteins whereas an excellent quadratic correlation (R2 = 0.9938) was observed between MADLS and SEC for the quantitative estimation of protein aggregation in the enzyme. Therefore, the approach provided using MADLS can be employed as a rapid screening method for the analysis of aberrations in different formulations and products prior to other Quality Control (QC) tests to speed up the batch release process. Overall, some of these alternative drying technologies offer a paradigm shift towards continuous manufacturing and allow controlled dry particle characteristics. The potential impact of these novel technologies can significantly reduce time, energy and costs associated with the manufacturing of biopharmaceuticals. The inclusion of Process Analytical Technology (PAT) and offline characterization techniques (described in Chapter 1), in tandem, provide additional information on the on the Critical Process Parameters (CPPs) and Critical Quality Attributes (CQAs) of biopharmaceutical products. Moreover, molecular modelling is a powerful tool that can reveal atomic-scale details to study the mechanisms of interactions of excipients with biologics. These technologies together can be envisaged to increase the manufacturing capacity of biopharmaceuticals at reduced costs as well as open avenues for further research and development.

Item Type: Thesis (Doctoral)
Uncontrolled Keywords: Biopharmaceuticals, Strategies
Departments or Groups: Pharmaceutical and Molecular Biotechnology Research Centre
Divisions: School of Science > Department of Chemical and Life Sciences
Depositing User: Derek Langford
Date Deposited: 26 Sep 2023 14:11
Last Modified: 26 Sep 2023 14:11

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