Development of a high-throughput platform for screening lipid nanoparticles for mRNA delivery

Abstract

mRNA lipid nanoparticles (LNPs) are at the forefront of nucleic acid intracellular delivery, as exemplified by the recent emergency approval of two mRNA LNP-based COVID-19 vaccines. The success of an...

Mechanistic Studies of an Automated Lipid Nanoparticle Reveal Critical Pharmaceutical Properties Associated with Enhanced mRNA Functional Delivery In Vitro and In Vivo

“…Previously, we established an automated high‐throughput platform for screening LNPs for mRNA delivery. [ 34 ] This automated high‐throughput platform is based on a common lab‐used liquid handling system and features fast and cost‐effective preparation, characterization, and in vitro evaluation of hundreds or thousands of mRNA LNPs. We have shown that LNPs made from the automated high‐throughput platform exhibited comparable or higher mRNA delivery efficiency than the LNPs generated from a standard microfluidic mixing platform.…”

“…[ 25,26,35 ] Indeed, using our automated high‐throughput mRNA LNP formulation technique, we demonstrated that a library of novel LNPs could be rapidly generated and screened compared to traditional microfluidics techniques. [ 34 ] We also surprisingly found that this automated technique imparted enhanced mRNA functional delivery over traditional microfluidics devices despite using the same lipid compositions. [ 34 ] In the current study, we sought to further investigate the mechanistic and physiochemical properties of LNPs prepared using this automated process.…”

“…[ 34 ] We also surprisingly found that this automated technique imparted enhanced mRNA functional delivery over traditional microfluidics devices despite using the same lipid compositions. [ 34 ] In the current study, we sought to further investigate the mechanistic and physiochemical properties of LNPs prepared using this automated process. Additionally, we explored LNP‐mediated mRNA delivery efficiency both in vitro and in vivo and found that LNPs formulated using the automated technique had a 4.5‐fold improvement in mRNA functional delivery in vivo as compared to LNPs formulated using traditional techniques.…”

“…To accelerate the study of LNPs, we previously developed an automated high‐throughput platform to integrate formulation preparation, biophysical characterization, and biological evaluation of LNPs encapsulating mRNA. [ 34 ] This platform enables generation of hundreds or thousands of LNPs in a time‐ and cost‐effective manner and can be applied to screen mRNA LNPs made from novel ionizable lipids in vitro. [ 34 ] It is acknowledged that the development of novel ionizable lipids with improved chemical structures represents a main delivery challenge as it requires the design of large lipid libraries which must be screened for delivery efficiency.…”

“…[ 34 ] This platform enables generation of hundreds or thousands of LNPs in a time‐ and cost‐effective manner and can be applied to screen mRNA LNPs made from novel ionizable lipids in vitro. [ 34 ] It is acknowledged that the development of novel ionizable lipids with improved chemical structures represents a main delivery challenge as it requires the design of large lipid libraries which must be screened for delivery efficiency. [ 25,26,35 ] Indeed, using our automated high‐throughput mRNA LNP formulation technique, we demonstrated that a library of novel LNPs could be rapidly generated and screened compared to traditional microfluidics techniques.…”