IP Library Granted Patent US 10,208,283
Granted Patent B2
US 10,208,283 · App. 15/000,749 · Granted Feb 19, 2019

Methods and systems for manipulating particles using a fluidized bed

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Quick Facts
Patent No.
US 10,208,283
App. No.
15/000,749
Granted
Feb 19, 2019
Kind
B2
Abstract

The present invention comprises methods and systems for manipulation of media and particles, whether inert materials or biomaterials, such as cells in suspension cell culture. The methods and systems comprise use of an apparatus comprising a rotating chamber wherein the actions of the combined forces of gravity, fluid flow force and centrifugal force form a fluidized bed within the rotating chamber.

Claims (64)

1. A method for manipulating particles using a fluidized bed, the method comprising:

rotating a chamber about a substantially horizontal axis to create a centrifugal force field, the chamber comprising an inlet and an outlet;

flowing a first stream containing a first media and particles into the chamber through the inlet, wherein flowing the first stream acts to create a force which opposes the centrifugal force;

forming a fluidized bed of particles in the chamber, wherein the forces substantially immobilize the particles in the fluidized bed by the summation of vector forces acting on the particles;

collecting the first media substantially without particles passing through the outlet of the chamber; then

manipulating the particles in the fluidized bed, wherein said manipulating is selected from the group consisting of removing, concentrating, diluting, exchanging media, harvesting, transferring, dispensing, transfecting, electroporating, separating, extracting, isolating, selecting, purifying, coating, binding, physically modifying, and altering the environment; and thereafter

removing the particles from the fluidized bed, wherein removing the particles comprises:

flowing a second stream into the chamber through the outlet, wherein flowing the second stream acts to create a force at least partially in the same direction as the centrifugal force field; and

collecting the particles passing through the inlet of the chamber.

2. The method of claim 1 , wherein the particles are cells, the method further comprising:

providing the first stream from a cell culture system prior to flowing the first stream into the chamber,

exchanging the media; and

delivering the cells and exchanged media to the cell culture system after removing the cells from the fluidized bed.

3. The method of claim 1 , wherein the particles are cells, wherein manipulating the cells comprises transfecting the cells, and wherein transfecting the cells comprises circulating a transfection stream containing a transfection reagent complex through the fluidized bed of cells one or more times.

4. The method of claim 1 , wherein the particles are cells, wherein manipulating the cells comprises electroporating the cells, and wherein electroporating the cells comprises:

applying an electric current to the fluidized bed of cells; and

altering the permeability of the cells.

5. The method of claim 4 , wherein electroporating the cells further comprises:

flowing a charged molecule stream containing charged molecules into the chamber through the inlet before, concurrently with, and/or after applying the electric current; and

incorporating the charged molecules into the cells.

6. The method of claim 1 , wherein manipulating the particles comprises concentrating the particles, wherein concentrating the particles comprises receiving the particles in a concentrated particles harvest container after removing the particles.

7. The method of claim 1 , wherein manipulating the particles comprises exchanging the media, and wherein exchanging the media comprises:

flowing a new media stream comprising a second media into the chamber through the inlet; and

replacing at least some of the first media in the fluidized bed with the second media.

8. The method of claim 1 , wherein manipulating the particles comprises harvesting the particles, and wherein harvesting the particles comprises receiving the particles in a particle harvest container after removing the particles.

9. The method of claim 1 , wherein manipulating the particles comprises dispensing the particles, and wherein dispensing the particles comprises receiving a measured amount of particles in one or more dispensed particle containers after removing the particles.

10. The method of claim 1 , wherein the particles comprise a mixed population of particles, wherein manipulating the particles comprises separating the mixed population of particles, and wherein separating the mixed population of particles comprises:

removing at least some of the mixed population of particles from the fluidized bed; and

collecting the at least some of the mixed population of particles passing through the outlet of the chamber.

11. The method of claim 10 , wherein removing at least some of the mixed population of particles comprises altering the centrifugal force field and/or the force of the first stream.

12. The method of claim 10 , wherein the mixed population of particles are separated by size, density, and/or shape.

13. The method of claim 1 , wherein manipulating the particles comprises coating the particles, and wherein coating the particles comprises:

flowing a coating stream containing a coating material into the chamber through the inlet; and

coating the particles retained in the fluidized bed with the coating material.

14. A method for separating a mixed population of particles, the method comprising:

rotating a chamber about a substantially horizontal axis, the chamber having an inlet and an outlet;

substantially immobilizing an affinity matrix in the chamber by forming a fluidized bed of the affinity matrix in the chamber;

flowing a first stream containing a first media and a mixed population of particles comprising target particles and non-target particles into the chamber through the inlet;

retaining target particles in the affinity matrix in the chamber; and

collecting the first media and non-target particles passing through the outlet of the chamber.

15. The method of claim 14 , further comprising:

flowing a second stream containing an elution media into the chamber through the inlet;

releasing the target particles from the affinity matrix; and

collecting the target particles passing through the outlet of the chamber.

16. A method for fractionating biomaterials, the method comprising:

rotating a chamber about a substantially horizontal axis to create a centrifugal force field, the chamber having an inlet and an outlet;

flowing a first stream containing a first media and a mixture of biomaterials into the chamber through the inlet, wherein flowing the first stream acts to create a force which opposes the centrifugal force;

selectively precipitating biomaterials from the first stream;

forming a fluidized bed of the precipitated biomaterials in the chamber, wherein the forces substantially immobilize the precipitated biomaterials in the fluidized bed by the summation of vector forces acting on the precipitated biomaterials; then

collecting the first media and the non-precipitated biomaterials passing through the outlet of the chamber; and thereafter

removing the precipitated biomaterials from the fluidized bed, wherein removing the precipitated biomaterials comprises:

flowing a second stream into the chamber through the outlet, wherein flowing the second stream acts to create a force at least partially in the same direction as the centrifugal force field; and

collecting the precipitated biomaterials passing through the inlet of the chamber.

17. The method of claim 16 , wherein the biomaterial is protein.

18. A method for separating a mixed population of particles, the method comprising:

rotating a chamber about a substantially horizontal axis, the chamber having an inlet and an outlet;

substantially immobilizing an affinity matrix in the chamber;

flowing a first stream containing a first media and a mixed population of particles comprising target particles and non-target particles from a separate non-rotating container into the chamber through the inlet while the chamber is rotating;

retaining target particles in the affinity matrix in the chamber; and

collecting the first media and non-target particles passing through the outlet of the chamber.

19. The method of claim 18 , further comprising:

flowing a second stream containing an elution media into the chamber through the inlet while the chamber is rotating;

releasing the target particles from the affinity matrix; and

collecting the target particles passing through the outlet of the chamber.

Assignments (2)
MERGER Recorded Jan 13, 2017
From: KSEP SYSTEMS, LLC
To: KSEP HOLDINGS, INC.
Reel/Frame 040966/0061 →
MERGER Recorded Jan 13, 2017
From: KSEP HOLDINGS, INC.
To: SARTORIUS STEDIM NORTH AMERICA INC.
Reel/Frame 040973/0480 →
Cited By (1)
US 12,453,976