The NanoAssemblr Platform: Microfluidics-based Manufacture of Limit Size Lipid Nanoparticles for Drug and Nucleic Acid Delivery

Colin L. Walsh1, Nathan M. Belliveau1,2, Paulo J.C. Lin1, Igor V. Zhigaltsev1, Justin B. Lee1, Robert J. Taylor2, Euan C. Ramsay2, Timothy J. Leaver2, Andre W. Wild1, Kevin Ou2, Alex K.K. Leung1, Yuen Yi C Tam1, Ying K. Tam4, Ismail Hafez1, Sam Chen1, Carl L. Hansen3, Pieter R. Cullis1

1 Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6Z 1Z3, Canada. 2 Precision NanoSystems, Inc., Vancouver, BC, V6Z 1Z3, Canada; 3 Department of Physics and Astronomy, University of British Columbia, Vancouver, BC, V6Z 1Z3, Canada. 4 Alcana Technologies, Vancouver, BC, V6Z 1Z3, Canada

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Summary
Here, we describe the NanoAssemblr platform: a novel, scalable microfluidics-based system for the development and manufacture of nanoparticles for drug delivery applications. The NanoAssemblr uses custom engineered microfluidic chips to enable millisecond mixing of nanoliter reaction volumes containing nanoparticle components. This well-controlled process mediates bottom-up self-assembly of nanoparticles with process and formulation dependent characteristics, including limit-sized lipid nanoparticles (lsLNP), the smallest stable structure a LNP can form. Rapid mixing provides precise control over nanoparticle formation, resulting in nucleic acid solid-core nanoparticles and bilayer vesicles with reproducible sizes (15-200 nm) and low polydispersity (PDI < 0.1). Additionally, this system is easily scaled by parallelizing microfluidic mixers, which maintains identical reaction conditions for batch sizes from milliliters to liters.

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