Methods for manufacturing an adjuvant
The present invention relates to compositions and methods for manufacturing an adjuvant comprising a saponin using a microfluidic device and to aspects thereof.
1. A solution comprising a solvent and 100-170 mg/ml phosphatidylcholine lipid, wherein the solvent comprises 70-90% v/v ethanol and 10-30% v/v isopropyl alcohol.
2. The solution according to claim 1 , comprising 100-160 mg/ml phosphatidylcholine.
3. The solution according to claim 2 , comprising 120-140 mg/ml phosphatidylcholine.
4. The solution according to claim 1 , further comprising 20-50 mg/ml sterol.
5. The solution according to claim 4 , comprising 30-40 mg/ml of sterol.
6. The solution according to claim 4 , wherein the ratio of phosphatidylcholine to sterol is 3:1 to 5:1.
7. The solution according to claim 4 , wherein the sterol is cholesterol.
8. The solution according to claim 1 , further comprising a Toll-like receptor 4 (TLR4) agonist.
9. The solution according to claim 8 , wherein the TLR4 agonist is a lipopolysaccharide.
10. The solution according to claim 9 , wherein the lipopolysaccharide is 3-de-O-acylated monophosphoryl lipid A (3D-MPL).
11. The solution according to claim 8 , wherein the TLR4 agonist is present at a concentration of 4-12 mg/ml.
12. The solution according to claim 1 , wherein the solvent comprises 75-85% v/v ethanol.
13. The solution according to claim 1 , wherein the solvent comprises 15-25% v/v isopropanol.
14. The solution according to claim 1 , wherein the solvent comprises 78-82% v/v ethanol, and 18-22% v/v isopropyl alcohol.
15. The solution according to claim 1 , wherein the phosphatidylcholine lipid contains saturated unbranched acyl chains having 12-20 carbon atoms.
16. The solution according to claim 15 , wherein the lipid is dioleoyl phosphatidylcholine (DOPC).
17. A solution which consists essentially of a solvent and 100-160 mg/ml DOPC, 30-40 mg/ml cholesterol, 4-10 mg/ml TLR4 agonist, and wherein the solvent comprises 70-90% v/v ethanol and 10-30% v/v isopropyl alcohol.
18. The solution according to claim 17 wherein the TLR4 agonist is a lipopolysaccharide.
19. The solution according to claim 18 , wherein the lipopolysaccharide is 3D-MPL.