Kinetic Control in Assembly of Plasmid DNA/Polycation Complex Nanoparticles

Yizong Hu, Zhiyu He, Yue Hao, Like Gong, Marion Pang, Gregory P. Howard, Hye Hyun Ahn, Mary Brummet, Kuntao Chen, Heng Wen Liu, Xiyu Ke, Jinchang Zhu, Caleb F. Anderson, Honggang Cui, Christopher G. Ullman, Christine A. Carrington, Martin G. Pomper, Jung Hee Seo, Rajat Mittal, Il MinnHai Quan Mao

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


Polyelectrolyte complex (PEC) nanoparticles assembled from plasmid DNA (pDNA) and polycations such as linear polyethylenimine (lPEI) represent a major nonviral delivery vehicle for gene therapy tested thus far. Efforts to control the size, shape, and surface properties of pDNA/polycation nanoparticles have been primarily focused on fine-tuning the molecular structures of the polycationic carriers and on assembly conditions such as medium polarity, pH, and temperature. However, reproducible production of these nanoparticles hinges on the ability to control the assembly kinetics, given the nonequilibrium nature of the assembly process and nanoparticle composition. Here we adopt a kinetically controlled mixing process, termed flash nanocomplexation (FNC), that accelerates the mixing of pDNA solution with polycation lPEI solution to match the PEC assembly kinetics through turbulent mixing in a microchamber. This achieves explicit control of the kinetic conditions for pDNA/lPEI nanoparticle assembly, as demonstrated by the tunability of nanoparticle size, composition, and pDNA payload. Through a combined experimental and simulation approach, we prepared pDNA/lPEI nanoparticles having an average of 1.3 to 21.8 copies of pDNA per nanoparticle and average size of 35 to 130 nm in a more uniform and scalable manner than bulk mixing methods. Using these nanoparticles with defined compositions and sizes, we showed the correlation of pDNA payload and nanoparticle formulation composition with the transfection efficiencies and toxicity in vivo. These nanoparticles exhibited long-term stability at -20 °C for at least 9 months in a lyophilized formulation, validating scalable manufacture of an off-the-shelf nanoparticle product with well-defined characteristics as a gene medicine.

Original languageEnglish (US)
Pages (from-to)10161-10178
Number of pages18
JournalACS Nano
Issue number9
StatePublished - Apr 30 2019


  • DNA/polycation nanoparticle
  • gene delivery
  • kinetic control
  • linear polyethylenimine
  • polyelectrolyte complex
  • transfection
  • turbulent mixing

ASJC Scopus subject areas

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)


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