Table 3. Examples of loading cubosomes (C) and bulk (B) cubic phases with proteins or oligonucleotides.

Examples of cubosomes loaded with proteins in order of increasing MW
Loaded active compound	MW (kDa)	Format (C/B)	Formulation	Reported observations	Ref.
Cholera toxin B subunit	12	C	Phy, glycolipid monosialoganglioside GM1, F127 stabilizer	Specific binding of cholera toxin B, indicating that cubosome systems could be developed for biosensing applications	[145]
Nerve growth factor	13.5	C	MO, beta casein stabilizer	Increased bioavailability vs free drug, measured in a guinea pig model	[115]
Human recombinant brain-derived neurotrophic factor (BDNF)	13.5	C	MO, eicosapentaenoic acid, PEG stabilizer	Time-resolved SAXS studying the phase transitions and kinetics of neurotrophin binding	[110]
Beta casein	24	C	MO or Phy and F127 stabilizer	Beta casein acts as a co-stabilizer and encourages formations of the hexagonal phase	[61]
Thermomyces lanuginosus lipase	32	C	Monoolein, F127 stabilizer	Enzymatic degradation of cubosomes	[116]
Outer membrane protein F (OmpF)	38	C	Monolinolein, octyl-POE stabilizer	Membrane protein allows interconnectivity between the two interconnected water channels, and can be controlled with pH	[114]
Ovalbumin	44.3	C	MO or Phy, F127 stabilizer	Encapsulation of ovalbumin with a sustained release profile	[41]
Ovalbumin	44.3	C	MO or Phy, monophosphoryl lipid A, imiquimod, F127 stabilizer	Cubosomes more efficient than liposomes at inducing an antigen specific cellular response in mice in vivo	[44]
Dopamine D2Lreceptor (membrane protein)	50	C	Ni(II) chelated EDTA amphiphiles	Amphiphiles allow increased loading of membrane proteins	[113]
Examples of cubosomes loaded with oligonucleotides
Loaded active compound	MW (kDa)	Format (C/B)	Formulation	Reported observations	Ref.
siRNA	19 base pairs	C	MO, DOTAP	Specific gene silencing with improved endosomal escape in comparison to liposomes	[91]
siRNA	19 base pairs	C	MO, DOTAP, MO-PEG	Gene knockdown in HeLa cells comparable to Lipofectamine	[45]
siRNA	22 base pairs	C	Phy, DOTAP, F127 stabilizer	Quartz crystal microbalance and SAXS study of siRNA and cubosome interactions	[120]
siRNA	22 base pairs	C	MO, DOTAP or DDAB, F127 stabilizer	Hexosome formation on loading, gene silencing	[119]
Salmon sperm DNA	Unknown	C	MO, cationic surfactants diisobutylphenoxyethyl-dimethylbenzyl ammonium chloride (DEBDA) or PEG-15 Cocopolyamine (PCPA), F127 stabilizer	Successful transfection in human HepG2 cell lines although further optimisation was necessary to increase efficiency	[142]
Examples of protein loading in bulk cubic phases in order of increasing MW
Loaded active compound	MW (kDa)	Format (C/B)	Formulation	Reported observations	Ref.
Horse radish peroxidase	44	B	Dimodan U/J a commercial-grade of monoglyceride (≥90%), sucrose stearate (0 - 20%)	Activity dependent on water channel size	[90]
Microbial rhodopsins, GPCRs, Heme-Copper Oxidases, Photosynthetic proteins, Outer membrane proteins	<50	B	MO, cholesterol	Thought to aid the nucleation of protein crystals which can then be interrogated with X-rays to determine protein structure	[151]
Cholesterol oxidase, horse radish peroxidase, glucose oxidase	50, 44, 80	B	Monolinolein	Demonstrated as detection systems for viruses (HIV, Ebola), naked eye test for malaria infected blood	[96]
D-fructose dehydrogenase (FDH) from Gluconobacter industrius	140	B	Monolinolein	No change in activity as a function of water channel size, but increased stability was demonstrated over 5 days	[89]
Gloeobacter ligand-gated ion channel (GLIC)	174	B	Monoacylglycerol (MP), 5-10% of DSPG	First crystallisation of a membrane protein with large extracellular domains via electrostatic swelling of the cubic phase	[86]