Horseshoe lamination mixer (HLM) sets new standards in the production of monodisperse lipid nanoparticles.

Microfluidic mixers promise unique conditions for the controlled and continuous preparation of nanoparticles by antisolvent precipitation. Nanoparticles may enable encapsulation of drug or mRNA molecules in the form of carrier nanoparticles or can provide higher bioavailability in the form of drug nanoparticles. The ultimate goal in microfluidic approaches is the production of nanoparticles with narrow size distributions while avoiding contaminations and achieving sufficiently high throughput. To achieve this, a novel microfluidic precipitation device was developed and realized by two-photon polymerization: mixing elements were designed in such a way that the liquids undergo a repeated Smale horseshoe transformation resulting in an increased interfacial area and mixing times of less than 10 ms. These elements and an additional 3D flow focusing ensure that no organic phase is exposed to the channel walls. The integration of a fluidic shield layer in the flow focusing proved to be useful to delay the precipitation process until reaching a sufficient distance to the injection nozzle. Lipid nanoparticle preparation with different concentrations of castor oil or the hard fat Softisan® 100 were performed at different flow rates and mixing ratios with and without a shield layer. Flow rates of up to 800 μl min -1 and organic phase mixing ratios of up to 20% resulted in particle sizes ranging from 42 nm to 166 nm with polydispersity indices from 0.04 to 0.11, indicating very narrowly distributed, and in most cases even monodisperse, nanoparticles. The occurrence of fouling can be completely suppressed with this new type of mixing elements, as long as Dean vortices are prevented. Moreover, this parameter range in the horseshoe lamination mixer provided a stable and continuous process, which enables a scalable production.