Method for processing solutions of biomolecules
Apparatus for in-line liquid exchanging a biomolecule-containing liquid is provided. The apparatus comprises a means ( 3 ) for mixing at least two liquids comprising a multiple inlet flow-controller ( 2 ), the means for mixing also comprising an outlet in fluid connection with a tangential flow filtration device ( 1 ) configured in single-pass mode.
1. Apparatus for in-line liquid exchanging a biomolecule-containing liquid comprising a means for mixing comprising a multiple inlet flow-controller further comprising two or more variable flow inlet valves for mixing at least two liquids, the means for mixing also comprising an outlet in fluid connection with a tangential flow filtration device configured in single-pass mode, wherein retentate from the tangential flow filtration device is in fluid connection with a second means for mixing at least two liquids and the second means for mixing comprises an outlet in fluid connection with a second tangential flow filtration device configured in single-pass mode.
2. Apparatus according to claim 1 , wherein the variable flow inlet valves are intermittent flow valves.
3. Apparatus for liquid exchanging a biomolecule-containing liquid comprising:
a) a multiple inlet flow-controller comprising:
i) a first inlet for a first liquid medium comprising a biomolecule;
ii) at least a second inlet for a second liquid medium;
iii) an outlet in fluid connection with a tangential flow filtration device; and
b) a means for imparting flow of the liquids through the flow-controller and the tangential flow filtration device, the means for imparting flow comprising a pump located between the outlet of the multiple inlet flow-controller and the tangential flow filtration device.
4. Apparatus according to claim 3 , further comprising a restrictor downstream of the tangential flow filtration device.
5. Apparatus according to claim 3 , further comprising a second multiple inlet flow-controller comprising:
i) a first inlet in fluid connection with the retentate from the tangential flow filtration device;
ii) a second inlet for a third liquid medium; and
iii) an outlet in fluid connection with a second tangential flow filtration device.
6. Apparatus according to claim 5 , wherein the second multiple inlet flow-controller functions as a restrictor.
7. Apparatus according to claim 6 , wherein the second multiple inlet flow-controller further comprises two or more variable flow inlet valves.
8. A method for the preparation of a biomolecule, which comprises processing a liquid medium comprising the biomolecule by liquid exchange employing apparatus for in-line liquid exchanging a biomolecule-containing liquid comprising a means for mixing comprising a multiple inlet flow-controller further comprising two or more variable flow inlet valves for mixing at least two liquids, the means for mixing also comprising an outlet in fluid connection with a tangential flow filtration device configured in single-pass mode, wherein the variable flow inlet valves are cycled between a position achieving a first, relatively low flow rate wherein the liquid remains able to flow, or flow is prevented and at least a second, higher flow rate.
9. The method according to claim 8 , wherein at least 10 cycles are employed.
10. The method according to claim 9 , wherein the cycle frequency is less than 100 Hz.
11. The method according to claim 8 , wherein the processing comprises buffer exchange.
12. A process for the production of a biomolecule which comprises a method according to claim 8 .
13. The process according to claim 12 , wherein the variable flow inlet valves are cycled between a position achieving a first, relatively low flow rate wherein the liquid remains able to flow, or flow is prevented and at least a second, higher flow rate, at least 10 cycles are employed, and the cycle frequency is from 0.05 to 0.5 Hz.
14. A method for the preparation of a biomolecule, which comprises processing a liquid medium comprising the biomolecule by liquid exchange employing an apparatus according to claim 1 .
15. A method according to claim 14 , wherein the variable flow inlet valves are cycled between a position achieving a first, relatively low flow rate wherein the liquid remains able to flow, or flow is prevented and at least a second, higher flow rate.
16. The method according to claim 15 , wherein at least 10 cycles are employed.
17. The method according to claim 16 , wherein the cycle frequency is less than 100 Hz.
18. The method according to claim 14 , wherein the processing comprises buffer exchange.
19. A process for the production of a biomolecule which comprises a method according to claim 14 .
20. The process according to claim 19 , wherein the variable flow inlet valves are cycled between a position achieving a first, relatively low flow rate wherein the liquid remains able to flow, or flow is prevented and at least a second, higher flow rate, at least 10 cycles are employed, and the cycle frequency is from 0.05 to 0.5 Hz.
21. A method for the preparation of a biomolecule, which comprises processing a liquid medium comprising the biomolecule by liquid exchange employing apparatus for in-line liquid exchanging a biomolecule-containing liquid comprising a means for mixing comprising a multiple inlet flow-controller further comprising two or more variable flow inlet valves for mixing at least two liquids, the means for mixing also comprising an outlet in fluid connection with a tangential flow filtration device configured in single-pass mode, wherein the variable flow inlet valves are cycled between a position achieving a first, relatively low flow rate wherein the liquid remains able to flow, or flow is prevented and at least a second, higher flow rate, at least 10 cycles are employed, and the cycle frequency is from 0.05 to 05 Hz.
22. A method for the preparation of a biomolecule, which comprises processing a liquid medium comprising the biomolecule by liquid exchange employing an apparatus according to claim 3 .
23. A method for the preparation of a biomolecule, which comprises processing a liquid medium comprising the biomolecule by liquid exchange employing an apparatus according to claim 7 , wherein the variable flow inlet valves are cycled between a position achieving a first, relatively low flow rate wherein the liquid remains able to flow, or flow is prevented and at least a second, higher flow rate.
24. The method according to claim 23 , wherein at least 10 cycles are employed.
25. The method according to claim 24 , wherein the cycle frequency is less than 100 Hz.
26. A process for the production of a biomolecule which comprises a method according to claim 23 .
27. The process according to claim 26 , wherein at least 10 cycles are employed, and the cycle frequency is from 0.05 to 0.5 Hz.