Formulation of Peptide-Based Nanoparticles Using a Microfluidic Device

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Lipid nanoparticles (LNPs) are common for nucleic acid delivery but require strict formulation control.
• Peptide-based nanoparticles (PBNs) present a promising alternative with potentially simpler formulation processes.

Purpose of the Study:

• To evaluate the formulation of WRAP5-based PBNs using microfluidics.
• To assess the impact of process parameters (flow rate ratio, total flow rate, mixing channel) on PBN characteristics.
• To determine the stability and biological activity of WRAP5-PBNs for nucleic acid delivery.

Main Methods:

• Microfluidic device used for formulating 72 different WRAP5-PBN formulations.
• Dynamic light scattering (DLS) for nanoparticle size and polydispersity index (PdI) analysis.
• Stability studies at 4°C and biological assays (gene silencing and expression) in relevant cell lines.

Main Results:

• Consistent nanoparticle size (50-70 nm) and low PdI (<0.22) were achieved, irrespective of formulation parameters.
• siRNA-loaded PBNs showed moderate size increase during storage; pDNA-loaded PBNs remained stable for 70 days.
• Effective CDK4 gene silencing (~50%) and mCHERRY gene expression were confirmed in cellular assays.

Conclusions:

• WRAP5-based PBN formulation via microfluidics is robust and scalable.
• These PBNs exhibit consistent physicochemical properties and potent biological activity.
• WRAP5-PBNs present a viable and potentially less complex alternative to LNPs for nucleic acid delivery.