Lipid nanoparticles (LNPs) are vital for the successful delivery of mRNA-based vaccines, including those of Moderna and Pfizer/BioNTech against SARS-CoV-2. Beyond viral vaccines, mRNA therapeutics offer promising applications in neonatal gene therapy, immunotherapy, and protein replacement therapy. Understanding the molecular interactions that LNPs undergo upon injection into biological systems is central to optimize the outcomes of nanoparticle-based delivery biotechnologies in clinical settings.<br/><br/>In this work, we study nano-bio interactions encountered by LNP-based delivery vehicles in biological matrices by probing what is known as the “corona” of adsorbed proteins that spontaneously forms on the LNPs in biological fluids. Although protein corona formation has traditionally been studied on rigid model nanoparticles, we anticipate that soft, biodegradable nanoparticles will be the workhorse for nucleic acid delivery applications. Accordingly, we develop an experimental workflow for collecting corona proteins from LNPs and quantifying the resultant protein corona with proteomic-based mass spectrometry and structural characterization. Importantly, this workflow is readily translatable across soft nanotechnologies of interest, which are understudied due to the experimental complexity of separating nanoparticle-protein complexes from free proteins. We investigate how modifying (i) the mRNA packaged inside the LNPs and (ii) the ionizable lipid within the LNPs modulate the subsequently formed protein corona in (iii) various biological environments of relevance for delivery applications. This fundamental understanding of protein-LNP interactions could enable more seamless design and clinical application of next-generation LNP carriers, thus supporting the safe and effective delivery of mRNA and other therapeutics to patients.

Protein corona formation on lipid nanoparticle-based delivery vehicles

ACS Spring 2022

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