Microfluidic synthesis of PEI/plasmid DNA polyplexes for downstream recombinant AAV vector synthesis: Physico-chemical and in vitro transfection study

Eş İ.

Chemical Engineering Science, cilt.317, 2025 (SCI-Expanded, Scopus) identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 317
  • Basım Tarihi: 2025
  • Doi Numarası: 10.1016/j.ces.2025.122075
  • Dergi Adı: Chemical Engineering Science
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Chemical Abstracts Core, Chimica, Compendex, INSPEC, zbMATH
  • Anahtar Kelimeler: Adeno-associated virus, Gene therapy, Microfluidics, pDNA, Polyethylenimine, Polyplex
  • Ankara Üniversitesi Adresli: Hayır

Özet

Adeno-associated virus (AAV) vectors play a crucial role in gene therapy and offer precise gene delivery with minimal immunogenicity and long-term expression. Efficient production of such vectors requires the co-delivery of capsid, expression, and helper pDNA into the nucleus. The synthesized particles should possess uniform physicochemical properties to ensure consistent transfection efficiency. Microfluidic technology offers a streamlined approach for the production of such particles by enabling single-run synthesis. In this study, we used hydrodynamic flow focusing (HFM), 3D swirling micromixer, for the synthesis of PEI/pDNA polyplexes and optimizing the conditions for polyplex production. Physicochemical analysis of the synthesized polyplexes showed each polyplex produced in different micromixer showed difference. In vitro transfection studies on HEK293 cells demonstrated a varying transfection efficiency for each micromixer. These findings suggest that the micromixers are promising tools for cost-effective and continuous AAV vector production.