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. 2017 Sep 21;7(1):12045.
doi: 10.1038/s41598-017-11533-1.

Formation and purification of tailored liposomes for drug delivery using a module-based micro continuous-flow system

Nikolay Dimov  1 , Elisabeth Kastner  2 , Maryam Hussain  3 , Yvonne Perrie  3 , Nicolas Szita  4
Affiliations

Formation and purification of tailored liposomes for drug delivery using a module-based micro continuous-flow system

Nikolay Dimov et al. Sci Rep. .

Erratum in

Abstract

Liposomes are lipid based bilayer vesicles that can encapsulate, deliver and release low-soluble drugs and small molecules to a specific target site in the body. They are currently exploited in several nanomedicine formulations. However, their development and application is still limited by expensive and time-consuming process development and production methods. Therefore, to exploit these systems more effectively and support the rapid translation of new liposomal nanomedicines from bench to bedside, new cost-effective and scalable production methods are needed. We present a continuous process flow system for the preparation, modification and purification of liposomes which offers lab-on-chip scale production. The system was evaluated for a range of small vesicles (below 300 nm) varying in lipid composition, size and charge; it offers effective and rapid nanomedicine purification with high lipid recovery (> 98%) combined with effective removal of non-entrapped drug (propofol >95% reduction of non-entrapped drug present) or protein (ovalbumin >90% reduction of OVA present) and organic solvent (ethanol >95% reduction) in less than 4 minutes. The key advantages of using this bench-top, rapid, process development tool are the flexible operating conditions, interchangeable membranes and scalable high-throughput yields, thereby offering simultaneous manufacturing and purification of nanoparticles with tailored surface attributes.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Particle size and polydispersity as a function of increasing backpressures in the TFF system as collected on the retentate side of the membrane. Images from NTA analysis, verifying particles in permeate (top) and retentate (bottom) stream at increasing backpressures. Particles were found in the permeate at backpressures exceeding 75 psi. All experimental datasets are presented as mean and standard deviation (mean ± s.d.) resulting from three independent runs (n = 3).
Figure 2
Figure 2
(A) Vesicle size, polydispersity (PDI), zeta potential (ZP) and particle concentration (P/mL) for cationic (DDA:TDB) and anionic (DPPG:DPPC:Chol) liposomes before and after the TFF purification. (B) Images from NTA show vesicles present in the retentate side only. (C) Propofol and ethanol removal achieved over three diafiltration cycles for anionic liposomes (DPPG:DPPC:Chol), expressed as a percentage of the initial amount of contaminants present.
Figure 3
Figure 3
Vesicle size, polydispersity (PDI), zeta potential (ZP) and particle concentration (P/mL) for (A) anionic liposomes (DPPG:DPPC:Chol) and (B) cationic liposomes (DDA:TDB) prior and post OVA-addition (ovalbumin, 100 μg mL−1), and particle characteristics after the TFF purification. Protein (ovalbumin) and ethanol removal achieved over three diafiltration cycles for (C) anionic and (D) cationic liposomes, expressed as a percentage of the initial amount of contaminants present. All experimental datasets are presented as mean and standard deviation (mean ± s.d.) average of three independent runs (n = 3).
Figure 4
Figure 4
(A) Schematic overview of the module-based microfluidic system. Liposomes were manufactured with a Staggered Herringbone Mixer (SHM) upstream and flowed through the Tangential Flow Filtration (TFF) device for consecutive purification. (B) Schematic overview of the formation of liposomes loaded with a low-solubility model drug, i.e. propofol. Vesicle assembly and drug loading are performed with a SHM, and non-entrapped (free) drug is removed from the mixture by consecutive filtration inside the TFF system. (C) Schematic overview of the formation of liposomes loaded with a model protein, ovalbumin (OVA). Vesicle assembly is performed with a SHM, with post-assembly protein addition; non-entrapped (free) protein is removed by consecutive diafiltration cycles inside the TFF system.
Figure 5
Figure 5
Lipid recovery in the continuous liposome factory-on-a-bench for (A) lipid recovery after four diafiltration cycles. (B) Lipid concentration in four concentration cycles, related to the initial amount of lipids present prior to the concentration cycles. All experimental datasets are presented as mean and standard deviation (mean ± s.d.) average of three independent runs (n = 3).
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