Thomas Anitha, M Garg Shyam, De Souza Rebecca A G, Ouellet Eric, Tharmarajah Grace, Reichert Dave, Ordobadi Mina, Ip Shell, Ramsay Euan C
Precision NanoSystems, Vancouver, BC, Canada.
Methods Mol Biol. 2018;1792:193-203. doi: 10.1007/978-1-4939-7865-6_14.
Lipid nanoparticles (LNPs) are established in the biopharmaceutical industry for efficient encapsulation and cytosolic delivery of nucleic acids for potential therapeutics, with several formulations in clinical trials. The advantages of LNPs can also be applied in basic research and discovery with a microfluidic method of preparation now commercially available that allows preparations to be scaled down to quantities appropriate for cell culture. These preparations conserve expensive nucleic acids while maintaining the particle characteristics that have made LNPs successful in later stages of genetic medicine development. Additionally, this method and the resulting LNPs are seamlessly scalable to quantities appropriate for in vivo models and development of nucleic acid therapeutics.The present work describes the methodology for preparing LNPs loaded with siRNA, mRNA or plasmids using a commercially available microfluidic instrument and an accompanying transfection kit. Guidelines for application to cultured cells in a well-plate format are also provided.
脂质纳米颗粒（LNPs）在生物制药行业中已被确立用于高效封装核酸并将其胞质递送以用于潜在治疗，有几种制剂正在进行临床试验。LNPs的优势也可应用于基础研究和发现，现在有一种商业化的微流体制备方法，该方法能将制剂规模缩小至适合细胞培养的量。这些制剂既能节省昂贵的核酸，又能保持使LNPs在基因药物开发后期取得成功的颗粒特性。此外，这种方法及由此产生的LNPs可无缝扩大规模至适合体内模型和核酸治疗药物开发的量。本工作描述了使用市售微流控仪器和配套转染试剂盒制备负载小干扰RNA（siRNA）、信使核糖核酸（mRNA）或质粒的LNPs的方法。还提供了以微孔板形式应用于培养细胞的指南。
