(624a) Continuous Manufacturing of Model Nanoparticle Systems for Peptide-Based Nanoparticle Gene Delivery

A major challenge in effective gene therapy and vaccine development is the inherent immunogenicity and degradability of nucleic acids in the body. Non-viral drug delivery vehicles can efficiently encapsulate nucleic acids and enhance residence time in the body. The only clinically approved, non-viral nucleic acid delivery vehicles are lipid-based nanoparticles, most notably applied in the COVID-19 vaccine.¹ However, harmful off-target effects upon systemic administration can be significant.² In recent years, novel delivery vehicles have been developed based on peptide nanoparticles. The self-assembly and cargo binding of these peptides or polypeptides are driven by their amphiphilic hydrophobic and cationic properties. Peptides are recognized by the body and may be tuned to improve targeting in the human body and reduce systemic toxicity. Clinical trials are in progress using peptide-based nanoparticles for gene delivery.

At present, the majority of peptide nanoparticles are produced using batch methods that are challenging to replicate and have limited scalability. This work aims to utilize impinging jet mixers (IJMs) which were used for the scale up of billions of doses of mRNA-based lipid nanoparticles to defeat the COVID-19 pandemic.³ Sourcing suitable peptides and cargo for the formulation of peptide-based nanoparticles is challenging. As a first step, we selected well-established nanoparticle systems which imitate peptide-nucleic acid nanoparticle properties and assembly. Next, we used these surrogate systems to investigate critical process parameter effects on critical quality attributes. The impact of pH, buffer, and mixing effects on size distribution, surface charge, morphology, and stability were characterized using inline process analytical technology and offline techniques. All experiments were conducted using a fully automated system, requiring minimal operator intervention during long duration experiments. These results will inform future work demonstrating at-scale manufacturing of peptide-based nanoparticles for gene delivery.

(1) Mendes BB, Conniot J, Avital A, et al. Nanodelivery of nucleic acids. Nat Rev Methods Primers. 2022;2(1):1-21. doi:10.1038/s43586-022-00104-y

(2) Boettler T, Csernalabics B, Salié H, et al. SARS-CoV-2 vaccination can elicit a CD8 T-cell dominant hepatitis. Journal of Hepatology. 2022;77(3):653-659. doi:10.1016/j.jhep.2022.03.040

(3) Devos C, Mukherjee S, Inguva P, et al. Impinging jet mixers: A review of their mixing characteristics, performance considerations, and applications. AIChE Journal. 2025;71(1):e18595. doi:10.1002/aic.18595