Apparatus for efficient genetic modification of cells

Patent metadata

Field	Value
Publication number	US-12139697-B2
Application number	US-202217946948-A
Country	US
Kind code	B2
Filing date	Sep 16, 2022
Priority date	Sep 20, 2017
Publication date	Nov 12, 2024
Grant date	Nov 12, 2024

Abstract

A device for treatment of cells with particles is disclosed. The device includes a semi-permeable membrane positioned between two plates, the first plate defining a first flow chamber and comprising a port, a flow channel, a transverse port, and a transverse flow channel, the first flow chamber constructed and arranged to deliver fluid in a transverse direction along the first side of the semi-permeable membrane, the second plate defining a second flow chamber and comprising a port. A method for transducing cells is disclosed. The method includes introducing a fluid with cells and viral particles into a flow chamber adjacent a semi-permeable membrane such that the cells and the viral particles are substantially evenly distributed on the semi-permeable membrane. The method also includes introducing a recovery fluid to suspend the cells and the viral particles, and separating the cells from the viral particles. A method of activating cells is disclosed.

First claim

What is claimed is:

1. A device for treatment of cells with particles, the device comprising:
a semi-permeable membrane having a plurality of pores dimensioned to allow passage of a fluid and prevent passage of the cells and the particles;
a substrate material having a lower hydraulic resistance than the semi-permeable membrane, the substrate material constructed and arranged to give structural support to the semi-permeable membrane;
the semi-permeable membrane and the substrate material positioned between first and second plates,
the first plate defining a first flow chamber adjacent to a first side of the semi-permeable membrane and comprising a port configured to deliver the fluid to the first flow chamber, a flow channel extending between the port and the first flow chamber, a transverse port configured to discharge the fluid, and a transverse flow channel extending between the transverse port and the first flow chamber, the first flow chamber being constructed and arranged to deliver the fluid in a substantially transverse direction along the first side of the semi-permeable membrane; and
the second plate defining a second flow chamber adjacent to a second side of the semi-permeable membrane and comprising a port configured to discharge the fluid from the second flow chamber.

2. The device of claim 1 , wherein the particles are viral particles or activation particles.

3. The device of claim 1 , further comprising a recycle loop extending between the port of the first plate and the port of the second plate.

4. The device of claim 1 , wherein the substrate material is further constructed and arranged to create a structured surface on the first side of the semi-permeable membrane, such that a monolayer of the cells and the particles are deposited substantially evenly across a surface of the first side of the semi-permeable membrane.

5. The device of claim 4 , wherein a surface area of the first side of the semi-permeable membrane is selected to correlate with a number and size of the cells.

6. The device of claim 5 , wherein the surface area of the first side of the semi-permeable membrane is between about 30 mm 2 and about 250 mm 2 for every 1 million cells.

7. The device of claim 1 , wherein the first flow chamber has a height between about 0.2 mm and about 2.0 mm.

8. The device of claim 7 , wherein the first flow chamber has a height between about 1.4 mm and about 1.8 mm.

9. The device of claim 1 , wherein the semi-permeable membrane has an average pore size of between about 50% and about 25% of the average diameter of the particles.

10. The device of claim 1 , wherein the semi-permeable membrane has an average pore size of 50 nm or less.

11. The device of claim 10 , wherein the semi-permeable membrane comprises a hydrophilic material exhibiting low protein binding characteristics.

12. The device of claim 11 , wherein the semi-permeable membrane comprises a material selected to limit the membrane protein fouling rate to about 10 mmHg/min or less for a flowrate of up to 0.4 ml/min.

13. The device of claim 12 , wherein the semi-permeable membrane comprises polyethersulfone (PES).

14. The device of claim 10 , wherein the semi-permeable membrane comprises at least one of polyvinylidene fluoride (PVDF), polycarbonate (PC), nylon, polypropylene, and polyethersulfone (PES).

15. A system comprising the device for treatment of cells with particles of claim 1 and
a device for separating the cells from the particles comprising:
a semi-permeable membrane having a plurality of pores dimensioned to allow passage of the fluid and the particles and prevent passage of the cells;
the transverse port being fluidly connectable to a port configured to deliver the fluid to a first flow chamber of the device for separating the cells from the particles.

16. The system of claim 15 , wherein the semi-permeable membrane of the device for separating the cells from the particles has an average pore size of between about 50% and about 25% of the average diameter of the cells.

17. The system of claim 15 , wherein the semi-permeable membrane of the device for separating the cells from the particles has an average pore size of between about 200 nm and 5 μm.

18. A method comprising separating the cells from the viral particles with the system of claim 15 .

19. A method comprising transducing the cells with the device of claim 1 .

20. A method comprising activating the cells with the device of claim 1 .

Assignees

• Charles Stark Draper Laboratory Inc

Inventors

• Kotz Kenneth T
• Teece Bryan D
• Truslow James Gillett
• Moore Nathan Francis
• Borenstein Jeffrey T
• Tandon Vishal

Classifications

• C12M41/46

  of cellular or enzymatic activity or functionality, e.g. cell viability · CPC title

• C12N2750/10043

  viral genome or elements thereof as genetic vector · CPC title

• C12M41/12

  of temperature (controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00) · CPC title

• C12M21/18 Primary

  Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes · CPC title

• C12M27/18

  Flow directing inserts · CPC title