Flow-through paramagnetic particle-based cell separation and paramagnetic particle removal

Patent metadata

Field	Value
Publication number	US-11912978-B2
Application number	US-202117503186-A
Country	US
Kind code	B2
Filing date	Oct 15, 2021
Priority date	Jun 5, 2015
Publication date	Feb 27, 2024
Grant date	Feb 27, 2024

Abstract

The present disclosure relates to systems and methods for flow-through separation of paramagnetic particle-bound cells in a cell suspension containing both bound and unbound cells as well as systems and methods for removing paramagnetic particles from paramagnetic particle-bound cells or from a cell suspension with unbound cells. It further relates to a flow-through magnetic separation/debeading module and a flow-through spinning membrane debeading module.

First claim

The invention claimed is:

1. A cell processing system comprising:
a flow-through magnetic separation/deb eading module, comprising:
a flow chamber defined by a plurality of walls,
an inlet arranged at a first end of the flow chamber for flowing a cell suspension comprising paramagnetic particle-bound cells and unbound cells into the flow chamber,
an outlet arranged at a second end of the flow chamber opposite the first end of the flow chamber,
a magnet assembly adjacent or proximate a wall of the flow chamber and configured to assert a magnetic force within the flow chamber,
a plurality of fluid injection ports arranged along a wall of the plurality of walls of the flow chamber, the plurality of fluid injection ports being positioned between the first end and the second end of the flow chamber, and
a membrane located above the plurality of fluid injection ports, wherein the membrane is configured to allow a fluid introduced through the plurality of injection ports to supply a force to any cells of the cell suspension stuck to the membrane sufficient to dislodge the cells from the membrane;

wherein the magnetic separation/debeading module is configured to establish the magnetic force within the flow chamber and the flow of the cell suspension through the flow chamber to both separate the paramagnetic particle-bound cells from the unbound cells and to debead a portion of the paramagnetic particle-bound cells.

2. The cell processing system of claim 1 , further comprising a spinning membrane debeading module positioned downstream from the at least one flow-through magnetic separation/debeading module.

3. The cell processing system of claim 1 , wherein the magnet assembly comprises at least two permanent magnets housed on a movable platform.

4. The cell processing system of claim 3 , wherein the movable platform is configured to move to adjust an amount of the magnetic force asserted within the flow chamber.

5. The cell processing system of claim 1 , wherein the magnet assembly comprises an electromagnet.

6. The cell processing system of claim 1 , wherein the walls of the flow chamber are flexible.

7. The cell processing system of claim 1 , further comprising a return loop configured to return the separated unbound cells or the separated particle-bound cells back through the flow-through magnetic separation/debeading module.

8. The cell processing system of claim 7 , further comprising at least one additional module.

9. The cell processing system of claim 8 , wherein the at least one additional module comprises at least one physical separation module.

10. The cell processing system of claim 7 , wherein the at least one additional module comprises at least one media exchange module.

11. The cell processing system of claim 7 , wherein the at least one additional module comprises at least one cell concentration module.

12. The cell processing system of claim 7 , wherein the at least one additional module comprises at least one cell washing module.

13. A method for processing paramagnetic particle-bound cells, comprising:
flowing a cell suspension comprising paramagnetic particle-bound cells and unbound cells through a flow chamber of a flow-through magnetic separation/debeading module; and
flowing fluid through a plurality of fluid injection ports arranged along the flow-through magnetic separation/debeading module, the plurality of fluid injection ports being positioned between a first end and a second end of the flow-through magnetic separation/debeading module, the fluid flowing through the plurality of fluid injection ports dislodging cells from a membrane positioned adjacent to the plurality of fluid injection ports, the dislodged cells passing further through the flow chamber;
wherein the magnetic separation/debeading module establishes a magnetic force within the flow chamber and a flow of the cell suspension through the flow chamber to both separate the paramagnetic particle-bound cells from the unbound cells and to debead a portion of the paramagnetic particle-bound cells.

14. The method of claim 13 , further comprising:
flowing the separated paramagnetic particle-bound cells through the flow-through magnetic separation/debeading module again to debead the paramagnetic particle-bound cells using a return loop.

15. The method of claim 13 , wherein the flow chamber comprises a plurality of walls, wherein the flow-through magnetic separation/debeading module further comprises
a magnet assembly comprising at least two magnets arranged proximate at least one wall of the plurality of walls of the flow chamber.

16. The method of claim 13 , further comprising flowing the separated paramagnetic particle-bound cells through a spinning membrane debeading module.

17. The method of claim 13 , further comprising flowing the separated unbound cells through the flow-through magnetic separation/debeading module again using a return loop to separate any paramagnetic particle-bound cells from particle-bound cells that were not separated from the unbound cells.

Assignees

• Novartis Ag
• Univ Pennsylvania

Inventors

• Fachin Fabio
• Rietze Rodney
• Greene Michael R
• Cao Lan

Classifications

• C12M47/04 Primary

  Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04) · CPC title

• B03C1/01

  by addition of magnetic adjuvants · CPC title

• B03C1/0332 Primary

  using permanent magnets · CPC title

• B03C1/288

  disposed at the outer circumference of a recipient · CPC title

• B03C1/30

  Combinations with other devices, not otherwise provided for · CPC title