Fabrication of antigen-containing nanoparticles using microfluidics with Tesla structure

Area of Science:

• Biotechnology
• Nanotechnology
• Vaccine Development

Background:

• Polyethyleneimine (PEI)-based antigen delivery systems are crucial for therapeutic vaccines.
• Traditional bulk mixing methods for preparing PEI-based antigen nanoparticles suffer from wide size distribution and poor reproducibility.
• These limitations hinder the broader application of PEI-based antigen delivery systems.

Purpose of the Study:

• To develop an improved microfluidic method for fabricating PEI-based antigen-containing nanopolyplexes.
• To enhance the uniformity, reproducibility, and efficiency of nanopolyplex preparation.
• To evaluate the antigen cross-presentation efficiency of nanopolyplexes prepared using the novel method.

Main Methods:

• Development of a microfluidic chip with a Tesla structure for nanopolyplex preparation.
• Comparison of physicochemical parameters between nanopolyplexes prepared by bulk mixing, non-Tesla microfluidic chips, and Tesla structured microfluidic chips.
• Assessment of protein structure and bioactivity preservation during microfluidic processing.
• Evaluation of antigen cross-presentation efficiency using a bone marrow-derived dendritic cell (BMDC) model.

Main Results:

• The Tesla structured microfluidic method produced nanopolyplexes with significantly more uniform size distribution compared to bulk mixing and non-Tesla chips.
• Reproducibility of nanopolyplex preparation was markedly improved.
• The microfluidic method demonstrated robustness and reliability, with minimal influence from operating parameters.
• Nanopolyplexes prepared at an optimal weight ratio showed higher antigen cross-presentation efficiency than free antigens in the BMDC model.

Conclusions:

• Tesla structured microfluidics provides a superior fabrication method for PEI-based antigen-containing nanopolyplexes compared to bulk mixing.
• This microfluidic approach overcomes limitations of traditional methods, enhancing uniformity and reproducibility.
• The developed method expands the potential of PEI-based antigen delivery systems for therapeutic vaccine applications.