title: Continuous Production of Discrete Plasmid DNA-Polycation Nanoparticles Using Flash Nanocomplexation.
abstract: Despite successful demonstration of linear polyethyleneimine (lPEI) as an effective carrier for a wide range of gene medicine, including DNA plasmids, small interfering RNAs, mRNAs, etc., and continuous improvement of the physical properties and biological performance of the polyelectrolyte complex nanoparticles prepared from lPEI and nucleic acids, there still exist major challenges to produce these nanocomplexes in a scalable manner, particularly for lPEI/DNA nanoparticles. This has significantly hindered the progress toward clinical translation of these nanoparticle-based gene medicine. Here the authors report a flash nanocomplexation (FNC) method that achieves continuous production of lPEI/plasmid DNA nanoparticles with narrow size distribution using a confined impinging jet device. The method involves the complex coacervation of negatively charged DNA plasmid and positive charged lPEI under rapid, highly dynamic, and homogeneous mixing conditions, producing polyelectrolyte complex nanoparticles with narrow distribution of particle size and shape. The average number of plasmid DNA packaged per nanoparticles and its distribution are similar between the FNC method and the small-scale batch mixing method. In addition, the nanoparticles prepared by these two methods exhibit similar cell transfection efficiency. These results confirm that FNC is an effective and scalable method that can produce well-controlled lPEI/plasmid DNA nanoparticles.
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Santos
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ren
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: John
authors.initials: J
authors.lastname: Vandermark
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Maani M
authors.initials: MM
authors.lastname: Archang
authors.affiliation: Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.|Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
authors.forename: John-Michael
authors.initials: JM
authors.lastname: Williford
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Heng-Wen
authors.initials: HW
authors.lastname: Liu
authors.affiliation: Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.|Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Lee
authors.affiliation: Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.|Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.|Department of Mechanical Engineering, Whiting School of Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Tza-Huei
authors.initials: TH
authors.lastname: Wang
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.|Translational Tissue Engineering Center and Whitaker Biomedical Engineering Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA.
authors.forename: Hai-Quan
authors.initials: HQ
authors.lastname: Mao
mesh_terms: D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology; D010957:Plasmids / Q000737:chemistry*; D011073:Polyamines / Q000737:chemistry*; D000071228:Polyelectrolytes; D011094:Polyethyleneimine / Q000737:chemistry
chemical_list: D004247:DNA; D011073:Polyamines; D000071228:Polyelectrolytes; D011094:Polyethyleneimine
keywords: DNA nanoparticles; flash nanocomplexation; gene delivery; linear PEI; scalable production