BACKGROUND: Oligonucleotides (ONs) have shown great promise as therapeutic agents for various diseases. It is necessary to provide a protocol for preparation of ON-loaded lipid nanoparticles (LNPs) in a reproducible manner on a laboratory scale. MATERIALS AND METHODS: A 3-inlet microfluidic (MF) chip-based device was used to synthesize LNPs at the lipid/ON ratio of 10/1 (w/w) and at flow rates ranging from 50 to 1100 μl/min. A series of LNPs containing either antisense oligodeoxyribonucleotide (AS-ODN) or small-interfering RNA (siRNA) were synthesized. Bulk mixing was used as control. RESULTS: The MF method was shown to be particularly useful for synthesis of LNPs loaded with AS-ODN. The optimal range of flow rates for AS-ODN LNPs was found to be 100 to 200 μl/min. MF synthesis produced LNPs with lower polydispersity values. However, the MF was less effective in preparing LNPs loaded with siRNA, which may have been due to greater rigidity of double-stranded siRNA comparing to single-stranded AS-ODN. CONCLUSION: MF technology is a simple, affordable and reproducible method for production of ON-LNPs.
BACKGROUND:Oligonucleotides (ONs) have shown great promise as therapeutic agents for various diseases. It is necessary to provide a protocol for preparation of ON-loaded lipid nanoparticles (LNPs) in a reproducible manner on a laboratory scale. MATERIALS AND METHODS: A 3-inlet microfluidic (MF) chip-based device was used to synthesize LNPs at the lipid/ON ratio of 10/1 (w/w) and at flow rates ranging from 50 to 1100 μl/min. A series of LNPs containing either antisense oligodeoxyribonucleotide (AS-ODN) or small-interfering RNA (siRNA) were synthesized. Bulk mixing was used as control. RESULTS: The MF method was shown to be particularly useful for synthesis of LNPs loaded with AS-ODN. The optimal range of flow rates for AS-ODN LNPs was found to be 100 to 200 μl/min. MF synthesis produced LNPs with lower polydispersity values. However, the MF was less effective in preparing LNPs loaded with siRNA, which may have been due to greater rigidity of double-stranded siRNA comparing to single-stranded AS-ODN. CONCLUSION: MF technology is a simple, affordable and reproducible method for production of ON-LNPs.
Authors: N Maurer; K F Wong; H Stark; L Louie; D McIntosh; T Wong; P Scherrer; S C Semple; P R Cullis Journal: Biophys J Date: 2001-05 Impact factor: 4.033
Authors: Chee Guan Koh; Xulang Zhang; Shujun Liu; Sharon Golan; Bo Yu; Xiaojuan Yang; Jingjiao Guan; Yan Jin; Yeshayahu Talmon; Natarajan Muthusamy; Kenneth K Chan; John C Byrd; Robert J Lee; Guido Marcucci; L James Lee Journal: J Control Release Date: 2009-08-28 Impact factor: 9.776
Authors: Rohit Karnik; Frank Gu; Pamela Basto; Christopher Cannizzaro; Lindsey Dean; William Kyei-Manu; Robert Langer; Omid C Farokhzad Journal: Nano Lett Date: 2008-07-26 Impact factor: 11.189