title: Microfluidic assembly of lipid-based oligonucleotide nanoparticles.
abstract: 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.
  
  
  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.
  
  
  CONCLUSIONS
  MF technology is a simple, affordable and reproducible method for production of ON-LNPs.
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Ohio State University, 1012 Smith Lab, 174 West 18th Ave, Columbus, OH 43210, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Weiming
authors.initials: W
authors.lastname: Xue
authors.forename: Yun
authors.initials: Y
authors.lastname: Wu
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Huang
authors.forename: L James
authors.initials: LJ
authors.lastname: Lee
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Lee
mesh_terms: D002460:Cell Line; D020868:Gene Silencing; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008156:Luciferases / Q000378:metabolism; D044085:Microfluidics / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis*; D034741:RNA, Small Interfering / Q000378:metabolism; D012212:Rheology; D013873:Thionucleotides / Q000138:chemical synthesis*
chemical_list: D008055:Lipids; D009841:Oligonucleotides; D034741:RNA, Small Interfering; D013873:Thionucleotides; C408162:oblimersen; D008156:Luciferases