Droplet-Based Electroporation for High-Throughput Biomolecular Cargo Loading of Extracellular Vesicles and Exosomes

Optimizes Cargo Loading a Sub-Cellular Scale, Enabling the Scaling of Drug Manufacturing

This electroporation platform enables the efficient, large-scale loading of biomolecular cargo into extracellular vesicles (EVs) and exosomes. Extracellular vesicles (EVs) are biocompatible, have fast cellular uptake, and can penetrate biological barriers, making them an emerging tool for delivering therapeutic agents to a patient’s body. However, loading the cargo molecules into EVs is currently highly challenging and inefficient due to their small size and scale, often with membrane fusion and aggregation.

Researchers at the University of Florida have developed a microfluidic, droplet-based electroporation system for efficient biomolecular cargo loading into extracellular vesicles (EVs) and exosomes, improving the efficiency and throughput of cargo loading and downstream drug delivery applications, as well as the compatibility with GMP manufacturing and scaling up.

Application

Droplet-based electroporation strategy for efficient and high-throughput loading of therapeutic cargo into extracellular vesicles and exosomes

Advantages

• Surrounds EVs/exosomes in droplets throughout the electroporation process, shielding them from contamination and heat degradation
• Manipulates EVs/exosomes in a microfluidic channel, enabling precise control of electrotransfection
• Provides a continuous flow of generation of droplets, electroporation, and harvesting, increasing throughput and processing on a scale of milliliters to liters of droplets
• Platform generates uniform electric field distribution, lowering the required voltage for improving electroporation efficiency
• The biomolecular cargo can be a target or therapeutic molecule, enabling the development of immunotherapy, gene therapy, vaccine, or regenerative medicine to treat diseases

Technology

This electroporation-based strategy for cargo loading into extracellular vesicles (EVs) or exosomes uses droplets containing EVs or exosomes and biomolecular cargo. The droplets flow through a microfluidic channel between two electrodes. The electrodes generate a uniformly distributed electric field across the channel, passing through multiple droplets at a time, increasing the permeability of the EVs/exosome membranes, and facilitating efficient cargo loading. The flow of droplets through the channel, electroporation, and harvesting is continuous, leading to large-scale production of loaded EVs primed for therapeutic delivery.