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

Patent metadata

Field	Value
Publication number	US-2016355777-A1
Application number	US-201615172415-A
Country	US
Kind code	A1
Filing date	Jun 3, 2016
Priority date	Jun 5, 2015
Publication date	Dec 8, 2016
Grant date	—

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

1 . A cell processing system comprising:
at least one cell suspension module; at least one buffer module; at least one flow-through magnetic separation/debeading module; at least one non-magnetic output module; and at least one magnetic output module.

2 . The cell processing system of claim 1 , further comprising at least one return loop returning upstream of at least one flow-through magnetic separation/debeading module.

3 . The cell processing system of claim 1 , comprising at least two flow-through magnetic separation/debeading modules in parallel.

4 . The cell processing system of claim 1 , comprising at least two flow-through magnetic separation/debeading modules in series.

5 . The cell processing system of claim 1 , further comprising at least one additional module.

6 . The cell processing system of claim 5 , wherein the at least one additional module comprises at least one spinning membrane debeading module.

7 . The cell processing system of claim 6 , comprising at least two spinning membrane debeading modules in parallel.

8 . The cell processing system of claim 6 , comprising at least two spinning membrane debeading modules in series.

9 . The cell processing system of claim 5 , wherein the at least one additional module comprises at least one physical separation module.

10 . The cell processing system of claim 9 , wherein the at least one additional module comprises at least one magnetic column module.

11 . The cell processing system of claim 5 , wherein the at least one additional module comprises at least one media exchange module.

12 . The cell processing system of claim 5 , wherein the at least one additional module comprises at least one cell concentration module.

13 . The cell processing system of claim 5 , wherein the at least one additional module comprises at least one cell washing module.

14 . The cell processing system of claim 1 , wherein the flow-through magnetic separation/debeading module comprises:
a chamber defined by walls and having an x-direction, a y-direction, and a z-direction; an inlet and an outlet arranged on opposite ends of the chamber in the y-direction; and at least two magnets adjacent or proximate a wall of the chamber and arranged to establish a zero gradient line within the chamber between the inlet and the outlet.

15 . The cell processing system of claim 6 , wherein the spinning membrane debeading module comprises:
a debeading chamber define partially by a cylindrical side-wall; a porous spinning membrane having an interior and oriented co-axially with the cylindrical side-wall; a sample inlet; a waste output module connected to the interior of the spinning membrane; and a cell output module connected to the debeading chamber.

16 . The cell processing system of claim 15 , wherein the spinning membrane debeading module further comprises at least one magnet adjacent or proximate to the cylindrical side-wall.

17 . A flow-through magnetic separation/debeading module comprising:
a chamber defined by walls and having an x-direction, a y-direction, and a z-direction; an inlet and an outlet arranged on opposite ends of the chamber in the y-direction; and at least two magnets adjacent or proximate a wall of the chamber and arranged to establish a zero gradient line within the chamber between the inlet and the outlet.

18 . The module of claim 17 , comprising at least two inlets and at least two outlets.

19 . The module of claim 17 , further comprising at least three magnets adjacent or proximate a wall of the chamber and arranged to establish at least two zero gradient lines within the chamber between the inlet and the outlet.

20 . The module of claim 17 , further comprising at least four magnets arranged in two arrays on opposite sides of the chamber in the z-direction.

21 . The module of claim 18 , further comprising at least four magnets arranged in two arrays on opposite side of the chamber in the z-direction and cross-oriented in the x-y plane from near one inlet to near one outlet on the opposite side of the chamber in the z-direction.

22 . The module of claim 17 , further comprising:
sub-membrane injection ports adjacent a wall of the chamber also adjacent at least two magnets; and a membrane adjacent the sub-membrane.

23 . A spinning membrane debeading module comprising:
a debeading chamber define partially by a cylindrical side-wall; a porous spinning membrane having an interior and oriented co-axially with the cylindrical side-wall; a sample inlet; a waste output module connected to the interior of the spinning membrane; a cell output module connected to the debeading chamber; and at least one magnet adjacent or proximate to the cylindrical side-wall.

24 . The spinning membrane debeading module of claim 23 , further comprising a reagent module.

25 . The spinning membrane debeading module of claim 23 , wherein the porous spinning membrane has a pore size greater than that of a particle to be debeaded and less than that of a cell to be debeaded.

26 . A method of flow-through cell processing comprising flowing a cell suspension comprising paramagnetic particle-bound cells through a flow-through magnetic separation/debeading module to produce an unbound cell product,
wherein the paramagnetic particle-bound cells continue to move in the flow-through magnetic separation/debeading module through the flowing step, and wherein the flow-through magnetic separation/debeading module comprises:
a flow chamber defined by walls through which the cell suspension flows; and
at least two magnets arranged adjacent or proximate at least one wall.

27 . The method of claim 26 , wherein the cell suspension is flowed laminarly through the flow-through magnetic separation/debeading module.

28 . The method of claim 26 , wherein the cell suspension further comprises unbound cells and flowing the cell suspension through the flow-through magnetic separation/debeading module separates the paramagnetic particle-bound cells and the unbound cells.

29 . The method of claim 28 , wherein the cell suspension further comprises free paramagnetic particles and flowing the cell suspension through the flow-through magnetic separation/debeading module separates the free paramagnetic particles and the unbound cells.

30 . The method of claim 28 , further comprising flowing the separated unbound cells through the flow-through magnetic separation/debeading module a second or subsequent time using a return loop.

31 . The method of claim 28 , further comprising flowing the separated paramagnetic particle-bound cells through the flow-through magnetic separation/debeading module a second or subsequent time using a return loop.

32 . The method of claim 31 , further comprising debeading the paramagnetic particle-bound cells in the flow-through magnetic separation/debeading module during the second or subsequent time to produce paramagnetic particles and debeaded, unbound cells.

33 . The method of claim 32 , further comprising flowing the produced paramagnetic particles and debeaded, unbound cells through the flow-through magnetic separation/debeading module a third or subsequent time to separate the paramagnetic particles and the debeaded, unbound cells.

Assignees

• Novartis Ag
• Univ Pennsylvania

Inventors

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

Classifications

• A61P35/02

  specific for leukemia · CPC title

• B03C2201/26

  for use in medical or biological applications · CPC title

• C07K16/2803

  against the immunoglobulin superfamily · CPC title

• B03C1/01

  by addition of magnetic adjuvants · CPC title

• C07K16/2809

  against the T-cell receptor (TcR)-CD3 complex · CPC title