Liposome synthesis

Synthesis of liposomes using microfluidics

Efficient liposome nanoparticle synthesis

High throughput, reproducibility and small size distribution

Easy system implementation

Unbox, set up and start your experiments right away

Straightforward scale up

Produce your liposomes at any volume (µL to L)

Liposome nanoparticle synthesis

The liposome nanoparticle synthesis pack can be customized to fit your specific needs. It contains at least two pumping channels to flow your two solutions – generally water and solvent – needed to perform the liposome synthesis inside your mixing chip. Liposomes can be synthesized using this liposome synthesis pack!

The microfluidic chip where the liposome synthesis occurs is a staggered herringbone chip with two inlets and induces chaotic mixing even at a microfluidic scale. The first inlet is used to provide the liposomes in ethanol, and the second one is used to bring the aqueous solution and possibly a hydrophile load that will be encapsulated inside the newly formed LNP, like siRNA, for example (see the application tab). At lower flow rates, other chips, such as coflowing ones, can be used to optimize your liposome production yield.

The production can be easily scaled up by increasing the volumes and flow rates and parallelizing several micromixers instead of one, thus increasing the system’s overall throughput while maintaining monodispersity and yield.

The stability and the speed of the reaction directly depend on the flow rates of each fluid in the microfluidic channel. The Elveflow OB1 MK4 flow controller creates the flow, and the flow rates are measured and regulated thanks to a flow sensor that makes a feedback loop for continuous flow control.

The combination of these instruments is the fastest and most precise microfluidic flow control available on the market, guaranteeing the best possible liposome monodispersity.

Pressure-driven flow control systems like the OB1 are more suited for liposome synthesis than peristaltic or syringe pumps because they are pulseless and can reach the desired flow rate in the chip way faster than other systems.

Liposome synthesis pack setup

This plug-and-play instrument LPN synthesis pack ensures a good compatibility between the different instruments, allowing the experiment to be started right away. The whole system is controlled by a single software and can be used for a wide variety of applications.

Contact our experts for a detailed protocol for microfluidic liposome synthesis.

A typical liposome pack contains:

An OB1 flow controller (Elveflow)
2 Microfluidic flow sensor (eg. Galileo)
Reservoirs
Chemical solutions to perform liposome synthesis if required: lipids, surfactants,…
One or several Fluidic 187 Herringbone mixer from microfluidic ChipShop (a flow focusing chip can also be used)
T-junctions/9 manifolds if production parallelizing
All necessary accessories: tubing, connectors, filters, etc…
Software (Galileo user interface)

Liposome synthesis is also possible using a flow-focusing method with a different microfluidic chip.

Microfluidics herringbone liposome nanoprecipitation principle

Ribonucleic acid (RNA) is a critical polymeric molecule for gene regulation and expression. RNA interference (RNAi) is a method that silences genes by using sequence-specific small interfering RNA (siRNA).

Based on their mRNA counterpart nucleotide sequences, siRNAs block the production of specific proteins [1-3]. Lipid nanoparticles (LNP) are the most commonly used system for delivering siRNA for therapeutic applications in vivo, such as anti-tumor agents or polyneuropathy treatment [4-5].

BioNTech/Pfizer’s BNT162b2 and Moderna’s mRNA-1273 vaccines also use lipid nanoparticles as vehicles for mRNA delivery into the cytoplasm of host cells, which leads to the production of COVID-19-Neutralizing antibodies [5]. Thus, lipid nanoparticle synthesis has played a critical role in developing COVID-19 vaccines and other nanomedicines and is considered very promising for developing new drug delivery systems [6].

Liposome nanoparticles can be used as cosmetic delivery systems for other applications, like solid lipid nanoparticles (SLN) [7].

The liposome nanoparticle synthesis pack includes staggered herringbone micromixers that create chaotic flows. They are the most commonly used microfluidic chips for LNP synthesis [8]. The structure generates chaotic advection, which can induce rapid mixing at Reynolds numbers smaller than 1 [9], thus allowing a better encapsulation efficiency than bulk methods [10].

References

Whitehead, K., Langer, R. & Anderson, D. Knocking down barriers: advances in siRNA delivery. Nat Rev Drug Discov 8, 129–138 (2009).
Belliveau N., Huft J., Lin P., Chen S., Leung A., Leaver T., Wild A., Lee J., Taylor R., Tam Y., Hansen C., Cullis P., Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA, Molecular Therapy – Nucleic Acids, 1, 2012, e37, 2162-2531
Allen, Theresa M., and Pieter R. Cullis. “Drug delivery systems: entering the mainstream.” Science 303.5665 (2004), 1818-1822.
Sasayama Y., Hasegawa M., Taguchi E., Kubota K., Kuboyama T., Naoi T., Yabuuchi H., Shimai N., Asano M., Tokunaga A., Ishii T., Enokizono J., 2019, In vivo activation of PEGylated long circulating lipid nanoparticle to achieve efficient siRNA delivery and target gene knock down in solid tumors. Journal of Controlled Release
Let’s talk about lipid nanoparticles. Nat Rev Mater 6, 99 (2021).
Thi, T.T.H.; Suys, E.J.A.; Lee, J.S.; Nguyen, D.H.; Park, K.D.; Truong, N.P. Lipid-Based Nanoparticles in the Clinic and Clinical Trials: From Cancer Nanomedicine to COVID-19 Vaccines. Vaccines 2021, 9, 359
Sylvia A Wissing; Rainer H Müller (2003). Cosmetic applications for solid lipid nanoparticles (SLN), 254(1), 65–68.
Maeki M., Kimura N., Sato Y., Harashima H., Tokeshi M., 2018, Advances in microfluidics for lipid nanoparticles and extracellular vesicles and applications in drug delivery systems. Advanced Drug Delivery Reviews
N-T. Nguyen, Z. Wu, Micromixers-a review, J. Micromech. Microeng. 15, 2005, R1-R16.
Leung A., Hafez I., Baoukina S., Belliveau S., Zhigaltsev I., Afshinmanesh E., Tieleman P., Hansen C., Hope M., and Cullis P., The Journal of Physical Chemistry C 2012 116 (34), 18440-18450

Why use microfluidics for liposome nanoparticle synthesis?

Liposome nanoparticles with a mean diameter between 20 nm and 100 nm have been synthesized using a system equivalent to the one that can be assembled using this pack [1-2]. Liposomes have also been synthesized with similar dimensions with improved reproducibility [3].

Microfluidic nanoprecipitation enables a good throughput, monodispersity with tunable particle size, and, generally, a better control of the nanoparticle synthesis. The encapsulation efficiency is also higher at the microfluidic scale.

One of the other substantial advantages of a microfluidic platform compared to a bulk method for liposome synthesis is that this process can be easily included as a single step in a more complex operation by adding more microfluidic instruments and chips.

Schematic of siRNA lipid nanoparticle synthesis — Schematic of LNP-siRNA formulation process employing the staggered herringbone micromixer. The lipid mixture in ethanol and siRNA in aqueous solution are pumped separately into the two inlets of the microfluidic mixing device. Herringbone structures induce chaotic advection of the laminar streams causing rapid mixing of the ethanol and aqueous phases and correspondingly rapid increases in the polarity experienced by the lipid solution. At a critical polarity the precipitates form as LNPs. Figure adapted from [2].

References

Belliveau N., Huft J., Lin P., Chen S., Leung A., Leaver T., Wild A., Lee J., Taylor R., Tam Y., Hansen C., Cullis P., Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA, Molecular Therapy – Nucleic Acids, 1, 2012, e37, 2162-2531.
Rungta, Ravi L., et al. “Lipid nanoparticle delivery of siRNA to silence neuronal gene expression in the brain.” Molecular therapy. Nucleic acids 2.12 (2013).
Kotouček, J., Hubatka, F., Mašek, J. et al. Preparation of nanoliposomes by microfluidic mixing in herring-bone channel and the role of membrane fluidity in liposomes formation. Sci Rep 10, 5595 (2020).

Customize your microfluidics liposome nanoparticle production

Contact our experts to answer any questions about this liposome nanoparticle synthesis pack and how it can match your specifications!

– Check our other Packs for various applications –

How can we help your experiment?

The packs applications are still under development, so we are not able to give you the tips or troubleshooting advice that we usually give in our user guides and application notes for the possible challenges you could face for a specific application experiment. That being said, we can always guarantee reliable and high-precision microfluidic flow control. Our microfluidic instruments are high-performance, versatile and user-friendly. Our experts will bring support and expertise during the setup and implementation of this sensor calibration pack.

Can a pack be customized based on my specific application?

Yes! Our experts will establish which instruments are best suited for your application, such as the type of flow sensor or the number of flow controller channels you need to perform your experiment. Contact us using the “talk to our experts” green button above.

Can I buy individual instruments?

You can order our instruments on the product section of our website.