Scalable mRNA and siRNA Lipid Nanoparticle Production Using a Parallelized Microfluidic Device
Scalable mRNA and siRNA Lipid Nanoparticle Production Using a Parallelized Microfluidic Device
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DOI：
10.1021/acs.nanolett.1c01353
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发表时间：
2021-06-30
期刊：
影响因子：
10.8
通讯作者：
Mitchell, Michael J.
中科院分区：
文献类型：
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作者：
Shepherd, Sarah J.;Warzecha, Claude C.;Mitchell, Michael J.
推进用于RNA治疗的脂质纳米颗粒（LNP）的主要挑战是开发可以在各种药物开发规模中可靠生产的制剂。微流体可以产生具有精确定义的性质的LNP，但受到缩放吞吐量的挑战的限制。为了解决这一挑战，我们提出了一个可扩展的，并行化的微流体装置（PMD），它包含了一个阵列的128个混合通道，同时操作。与单个微流体通道相比，PMD实现了> 100倍的生产速率，而没有牺牲微流体产生的LNP的典型的期望的LNP物理性质和效力。在小鼠中，我们显示出与常规混合相比，使用PMD产生的包封因子VII siRNA或编码RNA酶的mRNA的LNP的上级递送，肝基因沉默分别增加4倍和荧光素酶表达增加5倍。这些结果表明，这种PMD可以产生新兴临床应用所需的可扩展和可再现的LNP制剂，包括RNA治疗剂和疫苗。
A major challenge to advance lipid nanoparticles (LNPs) for RNA therapeutics is the development of formulations that can be produced reliably across the various scales of drug development. Microfluidics can generate LNPs with precisely defined properties, but have been limited by challenges in scaling throughput. To address this challenge, we present a scalable, parallelized microfluidic device (PMD) that incorporates an array of 128 mixing channels that operate simultaneously. The PMD achieves a >100x production rate compared to single microfluidic channels, without sacrificing desirable LNP physical properties and potency typical of microfluidic-generated LNPs. In mice, we show superior delivery of LNPs encapsulating either Factor VII siRNA or luciferase-encoding mRNA generated using a PMD compared to conventional mixing, with a 4-fold increase in hepatic gene silencing and 5-fold increase in luciferase expression, respectively. These results suggest that this PMD can generate scalable and reproducible LNP formulations needed for emerging clinical applications, including RNA therapeutics and vaccines.