Anionic lipids direct efficient microfluidic encapsulation of stable and functionally active proteins in lipid...

Area of Science:

• Biotechnology
• Nanomedicine
• Cell Biology

Background:

• Protein therapeutics face limitations due to poor cell membrane permeability.
• Existing delivery methods are restricted to cell surface or serum targets.
• Lipid nanoparticles offer a potential solution for intracellular delivery of polar molecules.

Purpose of the Study:

• To develop a method for efficient encapsulation and intracellular delivery of functional proteins using lipid nanoparticles.
• To investigate the association of amphoteric proteins with anionic lipids for nanoparticle assembly.
• To demonstrate the efficacy of Encapsulated Proteins in Lipid NanoParticles (EP-LNPs) for cellular uptake and protein functionality.

Main Methods:

• Utilized anionic lipids to facilitate protein association and microfluidic chip assembly of EP-LNPs.
• Encapsulated proteins with diverse molecular weights and isoelectric points.
• Assessed encapsulation efficiency, stability, cellular uptake in mammalian and fungal cells, and intracellular protein functionality.

Main Results:

• Achieved high encapsulation efficiencies (70-90%) for proteins in EP-LNPs at a 1:20 protein:lipid ratio.
• Demonstrated long-term stability of encapsulated proteins (several months).
• Showcased efficient cellular entry of EP-LNPs into mammalian and fungal cells, with functional intracellular proteins.

Conclusions:

• EP-LNPs technology enables efficient and functional intracellular delivery of proteins.
• This platform overcomes cell membrane barriers, expanding the scope of protein therapeutics.
• Potential applications include delivery of antibodies, enzymes, and other protein-based drugs.