Automated Preparation of Oligonucleotide-Loaded Lipid Nanoparticles Using Andrew+™

The progress in RNA-based therapies has seen significant advancements, including the approval of pioneering treatments like patisiran in 2018 and the recent development of COVID-19 vaccines using Lipid nanoparticle (LNP) formulations. Researchers have optimized the effectiveness of these therapies by leveraging electrostatic complexation of anionic RNA, utilizing a composition of cationic and ionizable lipids, combined with other lipids. High-throughput screening using robotic automation in a 96-well plate format has emerged as an efficient method, demonstrating results comparable to the current state-of-the-art methods.

To automate the LNP formulation for screening purposes, a protocol was established using the Andrew+ liquid handling platform. The method was developed to overcome challenges such as rapid evaporation of ethanolic lipid solutions. The automated method maintained consistent particle size and high mRNA encapsulation efficiency. The transfection efficiency into HepG2 cells was comparable to both manual pipetting and microfluidics based LNP preparation methods. Thus, the automated method using Andrew+ has shown promise in streamlining LNP formulation and advancing the development of mRNA-based therapies, offering high precision and repeatability for the high-throughput screening of various LNPs.