Microfluidic production of mRNA-loaded lipid nanoparticles for vaccine applications

Figures & data

Figure 1. Schematic representation of a pegylated lipid nanoparticle and a liposome.

Figure 2. Current pipeline of lipid nanoparticles (Data retrieved at 2022.08.19 from: pharmacircle).

Figure 3. Self-assembly of a LNP within a microfluidics device.

Figure 4. Common designs of microchannels systems. a) T-junction, b) hydrodynamic flow focusing, c) staggered herringbone, d) baffle and e) bifurcation.

Table 1. In vivo studies using mRNA-loaded LNPs obtained by microfluidics.

Disease	Lipid composition	Delivery route	Animal model	Particle size (nm)	EE (%)	Company	Ref.
Infectious disease vaccines
Influenza virus	Ionizable cationic:lipid: DSPC:chol: PEG-lipid (50:10:38.5:1.5 molar %)	id/im	mices, ferrets, NHPs, humans	80–100	>90%	Valera, A Moderna Venture	[101]
Influenza virus	DSPC:chol:PEG-lipid:GLA (9.83:38.5:1.5:50:0.17 molar %)	im/id	NHPs	80–100	>95%	Moderna Therapeutics	[102]
Influenza virus	Ionizable lipid:DSPC:chol: PEG-lipid (50:10:38.5:1.5 molar %)	im/id	NHPs	80–100	>95%	Moderna Therapeutics	[86]
Chikungunya virus	Ionizable cationic lipid: DSPC: chol: PEG-lipid (50:10:38.5:1.5 molar %)	iv	mice, NHPs	80–100	>90%	Moderna Therapeutics	[107]
Powassan virus	Ionizable lipid: structural lipid:sterol:PEG-lipid (50:10:38.5:1.5 molar %)	im	mice	80–100	>90%	Moderna Therapeutics	[87]
SARS-CoV-2	TT3:DOPE:chol: DMG-PEG2000 (20:30:40:0.75 molar %)	i.m	mice	-	-	Ohio State University	[88]
SARS-CoV-2	SM-102:PEG2000-DMG:DSPC:chol (50:1.5:10:38.5 molar %)	i.m	NHPs	80–100	>90%	Moderna Therapeutics	[89]
Zika virus	Ionizable lipid:DSPC: chol: PEG-lipid (50:10:38.5:1.5 molar %)	i.d	mice, NHPs	~80	-	University of Pennsylvania in collaboration with Acuitas Therapeutics and BioNTech	[105]
Zika virus	Ionizable cationic lipid:DSPC:chol: PEG-lipid (50:10:38.5:1.5 molar %)	i.m	mice	80–100	>90%	Valera, A Moderna Venture	[90]
Influenza virus	Ionizable lipid:DSPC:cholesterol:PEG lipid (50:10:38.5:1.5 molar %)	i.m	mice	50–142	69–100%	Moderna Therapeutics	[103]
HIV	Ionizable lipid:DSPC: chol:PEG-lipid (50:10:38.5:1.5 molar %)	i.v	mice	~80	-	University of Pennsylvania in collaboration with Acuitas Therapeutics	[106]
HIV	Ionizable lipid: DSPC:chol: PEG-lipid (50:10:38.5:1.5 molar %)	i.v	humanized mice	-	-	Upenn	[91]
Rabies virus	Ionizable lipid:DSPC: chol:PEG-lipid (50:10:38.5:1.5 molar %)	i.m	mice	-	-	CureVac AG	[104]
Human cytomegalovirus	Ionizable lipid:DSPC:chol: PEG-lipid (50:10:38.5:1.5 molar %)	i.m	mice, NHPs	80–100	>90%	Moderna Therapeutics	[92]
Dengue	Ionizable lipid:DSPC: chol:PEG-lipid (50:10:38.5:1.5 molar %)	i.m	mice	80–100	>80%	Moderna Therapeutics	[93]
Ebola	Ionizable lipid:DSPC:chol:PEG-lipid (50:10:38.5:1.5 molar %)	i.m	guinea pigs	-	-	Moderna Therapeutics	[94]
Cancer immunotherapy
Lymphoma	DOPE:Chol:PEG-lipid:ionizable lipid:Pam3	i.m	mice	99.45/91.69	74.94/77.77%	Ewha Womans University	[109]
Melanoma	cKK-E12: DOPE: chol:C14-PEG2000:sodium lauryl sulfate (15:26:40.5:2.5:16 molar %)	sc	mice	70–100	-	David H. Koch Institute for Integrative Cancer Research	[108]
Melanoma	A18: DOPE, chol:PEG-C14 (35:16:37.5:2.5 molar %)	sc	mice	-	-	Translate Bio	[95]
Protein-replacement therapies
Hemophilia B	DSPC:Chol:PEG-lipid:ATX (7:40:3:50 molar %)	i.v	mice	-	-	Arcturus Therapeutics	[96]
Cystic fibrosis	DSPC:Chol:DMG-PEG2000:Dlin-MC3-DMA (10:38.5:1.5:50 molar %)	inhalation	mice	104.2	>99%	Oregon State University	[111]
Friedreich´s ataxia	Dlin-MC3-DMA:DSPC:chol:DMG-PEG2000 (55:10:32.5:2.5 molar %)	i.th	mice	75–85	>95%	pfizer	[110]
Acute intermittent porphyria	DSPC:Chol:PEG-lipid: heptadecan-9-yl 8-((2-hydroxyethyl)(8-(nonyloxy)-8- oxooctyl)amino)octanoate (10:38.5:1.5:50 molar %)	i.v	mice,rabbits,NHPs	80–100	>90%	Moderna Therapeutics	[97]

Figure 5. mRNA-loaded LNPs endocytosis mechanism.

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