Single pot organic solvent-free thermocycling technology for siRNA-ionizable LNPs: a proof-of-concept approach for alternative to microfluidicsopen access
- Authors
- De, Anindita; Ko, Young Tag
- Issue Date
- Dec-2022
- Publisher
- TAYLOR & FRANCIS LTD
- Keywords
- Thermocycling technology; cooling-heating cycle; siRNA; ionizable LNPs; LNPs stability
- Citation
- DRUG DELIVERY, v.29, no.1, pp.2644 - 2657
- Journal Title
- DRUG DELIVERY
- Volume
- 29
- Number
- 1
- Start Page
- 2644
- End Page
- 2657
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/85623
- DOI
- 10.1080/10717544.2022.2108523
- ISSN
- 1071-7544
- Abstract
- Ionizable LNPs are the latest trend in nucleic acid delivery. Microfluidics technology has recently gained interest owing to its rapid mixing, production of nucleic acid-ionizable LNPs, and stability of nucleic acid inside the body. Industrial scale-up, nucleic acid-lipid long-term storage instability, and high production costs prompted scientists to seek alternate solutions to replace microfluidic technology. We proposed a single-pot, organic solvent-free thermocycling technology to efficiently and economically overcome most of the limitations of microfluidic technology. New thermocycling technology needs optimization of process parameters such as sonication duration, cooling-heating cycle, number of thermal cycles, and lipid:aqueous phase ratio to formulate precisely sized particles, effective nucleic acid encapsulation, and better shelf-life stability. Our research led to the formulation of siRNA-ionizable LNPs with particle sizes of 104.2 +/- 34.7 nm and PDI 0.111 +/- 0.109, with 83.3 +/- 4.1% siRNA encapsulation. Thermocycling siRNA-ionizable LNPs had comparable morphological structures with commercialized microfluidics ionizable LNPs imaged by TEM and cryo-TEM. When compared to microfluidics ionizable LNPs, thermocycling siRNA-ionizable LNPs had a longer shelf life at 4 degrees C. Our thermocycling technology showed an effective alternative to microfluidics technology in the production of nucleic acid-ionizable LNPs to meet global demand.
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