Methods, devices and systems for 3-stage filtration

Patent metadata

Field	Value
Publication number	US-11667672-B2
Application number	US-202117331963-A
Country	US
Kind code	B2
Filing date	May 27, 2021
Priority date	May 7, 2018
Publication date	Jun 6, 2023
Grant date	Jun 6, 2023

Abstract

The present disclosure pertains to filtration methods comprising: passing a first fluid that comprises cells, cell debris and targeted product produced by the cells through a first filter thereby separating the first fluid into a first retentate comprising cells and a first permeate comprising targeted product and cell debris; combining resin beads having affinity for targeted product with the first permeate to form a second fluid containing resin beads with bound target product and cell debris; passing the second fluid through a second filter thereby separating the second fluid into a second retentate comprising resin beads with bound target product and a second permeate comprising cell debris; combining an elution buffer with the second retentate to form a third fluid that comprises a mixture of resin beads and unbound targeted product; and passing the third fluid through a third filter thereby separating resin beads from targeted product.

First claim

The invention claimed is:

1. A system comprising:
a first filter configured to separate a first fluid comprising cells, cell debris and a target product-into a first retentate comprising the cells and a first permeate comprising the target product and cell debris;
a first mixing apparatus configured to combine the first permeate with resin beads having affinity for the target product to form a second fluid comprising resin beads with bound target product;
a second filter configured to separate the second fluid into a second retentate comprising the resin beads with bound target product and a second permeate comprising cell debris;
a second mixing apparatus configured to combine the second retentate with an elution buffer to form a third fluid comprising the resin beads and unbound target product; and
a third filter configured to separate the third fluid into a third retentate comprising the resin beads and a third permeate comprising the unbound target product.

2. The system of claim 1 , wherein the first filter has a mean pore size ranging from 1 μm to 8 μm.

3. The system of claim 1 , wherein the second filter has a mean pore size ranging from 2 μm to 30 μm.

4. The system of claim 1 , wherein the third filter has a mean pore size ranging from 2 μm to 30 μm.

5. The system of claim 1 , wherein the first, second and third filters are independently tangential flow filters.

6. The system of claim 5 , wherein the first, second, and third tangential flow filter each independently comprises hollow fibers having an inner lumen diameter ranging from 1 mm to 7 mm and a wall thickness of 1 mm to 8 mm.

7. The system of claim 1 , wherein the system operates in batch mode or in a continuous filtration mode.

8. The system of claim 7 , configured in a continuous filtration mode to receive the first fluid from a cell culture vessel and to return the first retentate to the cell culture vessel.

9. The system of claim 8 , configured for the first mixer to receive the second retentate.

10. The system of claim 9 , configured for the second mixer to receive the third retentate.

11. The system of claim 1 , wherein the target product is a protein.

12. The system of claim 1 , wherein the target product is a monoclonal antibody or a polyclonal antibody.

13. The system of claim 1 , wherein the resin beads are functionalized with one or more ligands selected from the group consisting of Protein A, Protein G, Protein A/G, and Protein L.

14. A system comprising
a first filter configured to separate a first fluid comprising cells, cell debris and a target product into a first retentate comprising the cells and a first permeate comprising the target product and cell debris;
a first mixing apparatus configured to combine the first permeate with resin beads having affinity for the target product to form a second fluid comprising the resin beads with bound target product;
a second filter configured to separate the second fluid into a second retentate comprising the resin beads with bound target product and a second permeate comprising cell debris;
a second mixing apparatus configured to combine the second retentate with a wash solution to form a third fluid comprising the resin beads with bound target product;
a third filter configured to separate the third fluid into a third retentate comprising the resin beads with bound target product and a second permeate comprising any remaining cell debris;
a third mixing apparatus configured to combine the third retentate with an elution buffer to form a fourth fluid comprising the resin beads and unbound target product,
a fourth filter configured to separate the fourth fluid into a fourth retentate comprising the resin beads and a final permeate comprising the unbound target product.

15. The system of claim 14 , wherein the system operates in batch mode or in a continuous filtration mode and is configured to receive the first fluid from a cell culture vessel and to return the first retentate to the cell culture vessel.

16. The system of claim 15 , wherein the first filter has a mean pore size ranging from 1 μm to 8 μm.

17. The system of claim 16 , wherein the second, third and fourth filters independently have a mean pore size ranging from 2 μm to 30 μm.

18. The system of claim 17 , wherein each of the first, second, third and fourth filter is a tangential flow filter and each independently comprises hollow fibers having an inner lumen diameter ranging from 1 mm to 7 mm and a wall thickness of 1 mm to 8 mm.

19. The system of claim 14 , wherein the target product is a protein.

20. The system of claim 14 , wherein the target product is a monoclonal antibody or a polyclonal antibody.

21. The system of claim 14 , wherein the resin beads are functionalized with one or more ligands selected from the group consisting of Protein A, Protein G, Protein A/G, and Protein L.

Assignees

• Repligen Corp

Inventors

• Bransby Michael

Classifications

• B01D2315/14

  Batch-systems · CPC title

• B01D2315/10

  Cross-flow filtration · CPC title

• B01D69/08

  Hollow fibre membranes (manufacture of hollow fibres D01D5/24, D01F1/08) · CPC title

• B01D15/3809

  of the antigen-antibody type, e.g. protein A, G or L chromatography · CPC title

• B01D2317/025

  Permeate series · CPC title