Liposomal apparatus and manufacturing method
View Patent ↗The present invention provides apparatus and processes for producing liposomes. By providing a buffer solution in a first reservoir, and a lipid solution in a second reservoir, continuously diluting the lipid solution with the buffer solution in a mixing chamber produces a liposome. The lipid solution preferably comprises an organic solvent, such as a lower alkanol.
1. An apparatus for producing a lipid vesicle encapsulating a nucleic acid within the lipid vesicle, said apparatus comprising:
a first reservoir for holding an aqueous solution including a nucleic acid;
a second reservoir for holding an organic lipid solution, wherein the lipids present in said organic lipid solution are solubilized in a lower alkanol at a concentration of about 75% v/v to 100% v/v; and
a pump mechanism configured to pump said aqueous solution and said organic lipid solution into a mixing environment at about equal flow rates;
wherein said organic lipid solution mixes with said aqueous solution in the mixing environment to instantaneously produce a lipid vesicle encapsulating the nucleic acid within the lipid vesicle by diluting said lower alkanol to a concentration of between 45% v/v to about 60% v/v; and
wherein the mixing environment includes a T-connector, wherein said aqueous solution and said organic lipid solution are introduced into the T-connector as opposing flows at about 180° relative to each other and mixed within the T-connector.
2. The apparatus of claim 1 , wherein said nucleic acid is selected from the group consisting of a plasmid, an antisense polynucleotide, a ribozyme, tRNA, snRNA, siRNA, and pre-condensed DNA.
3. The apparatus of claim 1 , wherein said nucleic acid comprises an siRNA.
4. The apparatus of claim 1 , wherein said lower alkanol is selected from the group consisting of methanol, ethanol, propanol, butanol, pentanol, isomers thereof, and combinations thereof.
5. The apparatus of claim 1 , wherein said lower alkanol comprises ethanol.
6. The apparatus of claim 1 , wherein the lipid vesicle is a liposome.
7. The apparatus of claim 1 , wherein one or both of the first and the second reservoirs are temperature controlled.
8. The apparatus of claim 1 , wherein one or both of the first and the second reservoirs include a jacketed stainless steel vessel with an overhead mixer.
9. The apparatus of claim 1 , wherein the T-connector is polypropylene or stainless steel.
10. The apparatus of claim 1 , wherein the lipids present in said organic lipid solution comprise a phospholipid, cholesterol, a PEG-lipid, and a cationic lipid.
11. The apparatus of claim 1 , wherein the apparatus further comprises a tangential flow ultrafiltration system.
12. The apparatus of claim 1 , wherein the lipids present in said organic lipid solution are solubilized in said lower alkanol at a concentration of 100% v/v.
13. The apparatus of claim 1 , wherein the pump mechanism comprises a peristaltic pump.