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QbD product development: rapid optimization and scale-up of PBAE-based siRNA delivery via DoE-guided microfluidics
Adrian P E Kromer1, Laetitia J M Eller1, David C Jürgens1
1Department of Pharmacy, Ludwig-Maximilians-Universität Munich Butenandtstrasse 5-13 Haus B 81377 Munich Germany.
RSC Pharmaceutics
|February 25, 2026
Summary
This study developed a robust workflow for creating RNA delivery nanoparticles using microfluidics. Optimized formulations show high gene silencing efficiency and stability, enabling scalable production.
Area of Science:
- Nanotechnology
- Biomaterials Science
- Gene Therapy
Background:
- Poly(β-amino ester) (PBAE) nanoparticles show promise for RNA delivery.
- Lack of clear design rules hinders optimization of PBAE formulations.
- Critical Quality Attributes (CQAs) and Critical Process Parameters (CPPs) require definition for consistent performance.
Purpose of the Study:
- To establish a Quality by Design (QbD)-guided workflow for developing microfluidically manufactured siRNA nanoparticles.
- To identify formulation parameters influencing nanoparticle physicochemical properties and RNA delivery performance.
- To enable rapid optimization, reliable scale-up, and clinically relevant performance of PBAE-based siRNA carriers.
Main Methods:
- Combined Quality by Design (QbD) and Design of Experiments (DoE) with high-throughput microfluidics.
- Utilized Response Surface Modeling to analyze formulation parameters and nanoparticle attributes.
- Evaluated polymer candidates based on intraparticle stability and gene silencing efficacy.
Main Results:
- High total flow rates (TFR ≥ 10), N/P ratios ≥10, and a 1:3 buffer:ethanol flow rate ratio (FRR) yielded smaller, stable nanoparticles.
- A polymer with balanced hydrophobic/hydrophilic side chains showed optimal stability and gene silencing.
- Flow rate ratio significantly impacted transfection efficiency and gene knockdown kinetics.
- The lead formulation achieved ~95% gene knockdown after two weeks of storage at 4 °C.
- Scale-up production confirmed the transferability of optimized CPPs and preserved CQA profiles.
Conclusions:
- A robust and scalable QbD-guided workflow for microfluidic siRNA nanoparticle development was established.
- The workflow enables rapid optimization and reliable scale-up of PBAE-based RNA delivery systems.
- Optimized formulations demonstrate clinically relevant performance and stability, paving the way for advanced gene therapies.
