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Controlled Nucleation of Lipid Nanoparticles

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ABSTRACT

Purpose

We describe a nucleation-based method which allows for the generation of monodisperse lipid nanoparticles over a range of diameters. Using a set of novel zwitterionic lipids and inverse phosphocholine lipids with pKas ranging from 2 to 5, we showed how the hydrodynamic diameter of lipid nanoparticles can be systematically manipulated over a 60 nm to 500 nm size range.

Method

Lipid nanoparticles were prepared by adding an anti-solvent, such as water, to the organic phase containing the lipid components. This led to super-saturation and the spontaneous formation of particles.

Results

The growth and final particle size was controlled by the ratio of the components in the ternary system: lipid, organic solvent and aqueous phase. Particles with diameter below 125 nm were formed under conditions where the super-saturation coefficient was between 2.3 and 20. PEG-lipid served as an efficient growth inhibitor except at very high and low lipid concentrations. Encapsulation efficiency of siRNA into lipid nanoparticles was shown to be pH-dependent and requires the protonation of the anionic carboxylate groups of the zwitterionic lipids, emphasizing the importance of electrostatic forces.

Conclusion

This process enables high encapsulation efficiency of nucleic acids and allows the size of lipid nanoparticles to be controlled.

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ACKNOWLEDGMENTS & DISCLOSURES

We are grateful for the financial support from the NIH (2R01EB003008-08) and Pfizer QB3. Juliane Nguyen is supported by the postdoctoral research fellowship from the Deutsche Forschungsgemeinschaft (DFG). Colin Walsh and Emily Perttu are supported by the NSF pre-doctoral fellowship. Michael Motion is supported by a fellowship of the NIH NIGMS. We acknowledge Charles Noble and Mark Hayes for their technical help with the HPLC.

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Authors and Affiliations

  1. Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, 94143, USA

    Juliane Nguyen, Colin L. Walsh, J. P. Michael Motion, Emily K. Perttu & Francis Szoka

  2. The UC Berkeley–UCSF Graduate Program in Bioengineering, University of California Berkeley, Berkeley, California, 94720, USA

    Colin L. Walsh, J. P. Michael Motion, Emily K. Perttu & Francis Szoka

  3. University of California, HSE 1145, 513 Parnassus Avenue, San Francisco, California, 94141-0912, USA

    Francis Szoka

Authors
  1. Juliane Nguyen
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  2. Colin L. Walsh
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  3. J. P. Michael Motion
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  4. Emily K. Perttu
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  5. Francis Szoka
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Corresponding author

Correspondence to Francis Szoka.

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Nguyen, J., Walsh, C.L., Motion, J.P.M. et al. Controlled Nucleation of Lipid Nanoparticles. Pharm Res 29, 2236–2248 (2012). https://doi.org/10.1007/s11095-012-0752-2

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  • DOI: https://doi.org/10.1007/s11095-012-0752-2

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