Stable compositions containing OMVs
View Patent ↗An OMV preparation comprises OMVs having a sufficiently positive or negative surface charge to substantially prevent aggregation.
1. A method of preventing aggregation of outer membrane vesicles (OMVs) comprising:
(i) obtaining a preparation of OMVs in a solution,
(ii) measuring the aggregation of the OMVs in said solution,
(iii) if the OMVs are aggregated, adjusting zeta potential of the OMVs in the solution, and returning to step (ii).
2. A method according to claim 1 , wherein the OMVs are aggregated when they have a mean diameter in excess of 200 nm.
3. A method according to claim 1 , wherein the zeta potential is adjusted by altering pH of the solution.
4. A method according to claim 1 , wherein the zeta potential is adjusted by altering the ionic strength of the solution.
5. A method according to claim 1 , further comprising a homogenisation step after step (iii).
6. A method according to claim 1 , wherein the preparation of OMVs is subsequently frozen.
7. A method according to claim 1 , wherein magnitude of the zeta potential is adjusted to be more negative than the starting value by −5 mV to −30 mV, −10 mV to −20 mV, −5 mV to −15 mV, −8 mV to −12 mV, or −10 mV.
8. A method according to claim 1 , wherein the solution is a buffer solution.
9. A method according to claim 1 , comprising adjusting the zeta potential of the OMVs in the solution to fall outside the range −15 mV to 15 mV.
10. A method according to claim 3 , wherein the pH is altered to be equal to or greater than pH 8.
11. A method according to claim 3 , wherein the pH is altered to be equal to or less than pH 6.
12. A method according to claim 4 , wherein the ionic strength of the solution is altered by the addition of ions to the solution.
13. A method according to claim 1 , wherein the zeta potential is adjusted by altering temperature of the solution.
14. A method according to claim 13 , wherein the zeta potential is adjusted by lowering the temperature of the solution.
15. A method of according to claim 1 , wherein the preparation is subsequently filtered through a filter membrane.
16. A method according to claim 15 , further comprising a homogenisation step prior to the filtering step.
17. A method according to claim 15 , wherein, subsequent to filtering through the filtering membrane, at least 90% of the OMVs have a mean diameter of less than 200 nanometres.
18. A method according to claim 15 , wherein, subsequent to filtering through the filtering membrane, at least 90% of the OMVs have a mean diameter of less than 180 nanometres.
19. A method according to claim 15 , wherein, subsequent to filtering through the filing filtering membrane, at least 90% of the OMVs have a mean diameter of less than 160 nanometres.
20. A method according to claim 15 , wherein the filter membrane has a pore size of from 0.2-2 microns.
21. A method according to claim 15 for sterilizing an OMV preparation, wherein the filter membrane has a pore size of about 0.2 microns.