Collection, genome editing, and washing of T-cell lymphocytes

Patent metadata

Field	Value
Publication number	US-12168087-B2
Application number	US-202016807254-A
Country	US
Kind code	B2
Filing date	Mar 3, 2020
Priority date	Mar 5, 2019
Publication date	Dec 17, 2024
Grant date	Dec 17, 2024

Abstract

Blood from a blood source is drawn into a fluid flow circuit. A mononuclear cell product is separated from the blood, followed by at least a portion of the mononuclear cell product being conveyed into an electroporation device without disconnecting the blood source from the fluid flow circuit. The electroporation device opens pores in a membrane of at least one of the cells of the mononuclear cell product to allow DNA material (which is added to the mononuclear cell product prior to electroporation) to enter and modify the genome of the cell. At least a portion of the modified mononuclear cell product is returned to the blood source. The mononuclear cell product may be washed prior to being conveyed into the electroporation device. The modified mononuclear cell product may be washed after exiting the electroporation device.

First claim

The invention claimed is:

1. A fluid processing system, comprising:
a first separation device configured to separate mononuclear cells and T-cell lymphocytes from blood;
a second separation device that is separate from the first separation device;
an electroporation device;
a pump assembly including a plurality of pumps; and
a controller configured to execute a mononuclear cell processing procedure to
actuate the pump assembly to convey blood from a blood source into the first separation device,
actuate the first separation device to separate mononuclear cells and T-cell lymphocytes from the blood as a mononuclear cell product,
actuate the pump assembly to convey at least a portion of the mononuclear cell product into the electroporation device to modify a genome of at least one of the cells of the mononuclear cell product so as to create a modified mononuclear cell product, and
actuate the pump assembly to convey at least a portion of the modified mononuclear cell product to the blood source, wherein
the second separation device is configured to wash the mononuclear cell product and to wash the modified mononuclear cell product, and
the controller is configured to selectively control the second separation device during a first version of the mononuclear cell processing procedure to wash the mononuclear cell product, to control the second separation device during a second version of the mononuclear cell processing procedure to wash the modified mononuclear cell product, and to control the second separation device during a third version of the mononuclear cell processing procedure to wash neither the mononuclear cell product nor the modified mononuclear cell product.

2. The fluid processing system of claim 1 , wherein the first separation device comprises a centrifuge.

3. The fluid processing system of claim 1 , wherein the controller is configured to actuate the second separation device to wash the mononuclear cell product prior to actuating the pump assembly to convey said at least a portion of the mononuclear cell product into the electroporation device and to actuate the second separation device to wash the modified mononuclear cell product after actuating the pump assembly to convey said at least a portion of the mononuclear cell product into the electroporation device.

4. The fluid processing system of claim 1 , wherein the controller is configured to actuate the pump assembly to collect and retain the modified mononuclear cell product for a holding time prior to conveying said at least a portion of the modified mononuclear cell product to the blood source.

5. The fluid processing system of claim 1 , wherein the controller is configured to actuate the pump assembly to convey said at least a portion of the modified mononuclear cell product to the blood source without first collecting and retaining the modified cell product for a holding time.

6. The fluid processing system of claim 1 , wherein the electroporation device is configured to execute multiple steps of an electroporation-based procedure, including addition of DNA material to the mononuclear cell product and electroporation.

7. The fluid processing system of claim 1 , wherein the electroporation device is configured to subject the mononuclear cell product to electroporation, but not configured to add DNA material to the mononuclear cell product.

8. The fluid processing system of claim 7 , wherein the controller is configured to actuate the pump assembly to add DNA material to the mononuclear cell product.

9. The fluid processing system of claim 8 , wherein the controller is configured to control the pump assembly to retain the mixture of the DNA material and the mononuclear cell product for a holding time prior to actuating the pump assembly to convey said at least a portion of the mononuclear cell product into the electroporation device.

10. The fluid processing system of claim 1 , wherein the second separation device comprises a spinning membrane separator drive unit.

11. The fluid processing system of claim 1 , further comprising an outlet sensor associated with the second separation device and configured to detect optical characteristics of a waste product exiting the second separation device.

Assignees

• Fenwal Inc

Inventors

• Min Kyungyoon
• Kusters Benjamin E

Classifications

• A61M1/36225

  with blood pumping means or components thereof · CPC title

• A61M1/362261

  at least one cassette surface or portion thereof being flexible, e.g. the cassette having a rigid base portion with preformed channels and being covered with a foil · CPC title

• A61M1/362265

  Details of valves · CPC title

• A61M1/36224

  with sensing means or components thereof · CPC title

• A61M1/362227

  the interface providing means for actuating on functional elements of the cassette, e.g. plungers · CPC title