Coupling Lipid Nanoparticle Structure and Automated Single‐Particle Composition Analysis to Design Phospholipase‐Responsive Nanocarriers

Abstract

Lipid nanoparticles (LNPs) are versatile structures with tunable physicochemical properties that are ideally suited as a platform for vaccine delivery and RNA therapeutics. A key barrier to LNP rational design is the inability to relate composition and structure to intracellular processing and function. Here Single Particle Automated Raman Trapping Analysis (SPARTA) is combined with small‐angle X‐ray and neutron scattering (SAXS/SANS) techniques to link LNP composition with internal structure and morphology and to monitor dynamic LNP–phospholipase D (PLD) interactions. This analysis demonstrates that PLD, a key intracellular trafficking mediator, can access the entire LNP lipid membrane to generate stable, anionic LNPs. PLD activity on vesicles with matched amounts of enzyme substrate is an order of magnitude lower, indicating that the LNP lipid membrane structure can be used to control enzyme interactions. This represents an opportunity to design enzyme‐responsive LNP solutions for stimuli‐responsive delivery and diseases where PLD is dysregulated.

Nanotechnology’s frontier in combatting infectious and inflammatory diseases: prevention and treatment

“… 72 – 75 LNPs may be designed for stimuli-triggered release, but accuracy in physiological conditions remains a challenge. 76 – 79 Research and optimization are ongoing to address these limitations and enhance lipid NP performance.…”

Innovative Approaches for Drug Discovery: Quantifying Drug Distribution and Response with Raman Imaging

“…The authors successfully demonstrated the advanced capabilities of SPARTA by analyzing different nanoparticle formulations, including liposomes and polymersomes, as well as the functionalization of polystyrene particles. Since its initial demonstration, SPARTA has been used to study a multitude of nanoparticle systems in recent years. − Saunders et al adapted SPARTA to distinguish cargo location and loading heterogeneity within nanoparticle populations . They generated a theoretical model and showed a positive, linear correlation between the encapsulating polymer and cargo mass for membrane loading and a much lower cubic relationship for core loading.…”

Lipid Nanoparticle (LNP) Delivery Carrier-Assisted Targeted Controlled Release mRNA Vaccines in Tumor Immunity

“…Lipid nanoparticles (LNPs), as nanoparticle carriers containing mRNA, play an important role in whole-body transport [ 57 , 58 , 59 , 60 ]. Their superior biocompatibility and efficient intracellular release mechanism make them an ideal drug delivery tool [ 61 , 62 , 63 ].…”