End-to-end cell therapy automation

Patent metadata

Field	Value
Publication number	US-2020071670-A1
Application number	US-201916668745-A
Country	US
Kind code	A1
Filing date	Oct 30, 2019
Priority date	Sep 1, 2017
Publication date	Mar 5, 2020
Grant date	—

Abstract

The present disclosure provides an automated method of producing genetically modified immune cells, including chimeric antigen receptor T (CAR T) cells, utilizing a fully-enclosed cell engineering system.

First claim

1 . A method for automated production of a genetically modified immune cell culture, the method performed by a cell engineering system, comprising:
(a) activating an immune cell culture with an activation reagent to produce an activated immune cell culture in a first chamber of the cell engineering system; (b) transducing the activated immune cell culture, the transducing comprising:
i. transferring via a first sterile, closed connection, the activated immune cell culture from the first chamber to an electroporation unit;
ii. electroporating the activated immune cell culture with a vector, to produce a transduced immune cell culture;
iii. transferring via a second sterile, closed connection, the transduced immune cell culture to a second chamber of the cell engineering system;
(c) expanding the transduced immune cell culture; (d) concentrating the expanded immune cell culture of (c); and (e) harvesting the concentrated immune cell culture of (d) to produce a genetically modified cell culture.

2 . The method of claim 1 , wherein the electroporation unit is located outside of the cell engineering system.

3 . The method of claim 1 , wherein the method produces at least about 100 million viable genetically modified immune cells.

4 . The method of claim 1 , wherein the method produces at least about 2 billion viable genetically modified immune cells.

5 . The method of claim 1 , wherein the immune cell culture is a T cell culture.

6 . The method of claim 5 , wherein T cell culture is a chimeric antigen receptor T (CAR T) cell culture.

7 . The method of claim 6 , wherein the vector encodes a chimeric antigen receptor.

8 . The method of claim 1 , wherein the immune cell culture comprises peripheral blood mononuclear cells and/or purified T cells.

9 . The method of claim 1 , wherein the cell culture comprises at least one accessory cell.

10 . The method of claim 9 , wherein the accessory cell comprises a monocyte or a monocyte-derived cell.

11 . The method of claim 10 , wherein the accessory cell comprises antigens for a T cell receptor, including CD28, CD40, CD40L and/or ICOS.

12 . The method of claim 1 , wherein the activation reagent comprises an antibody or a dendritic cell.

13 . The method of claim 12 , wherein the antibody is immobilized on a surface.

14 . The method of claim 13 , wherein the surface is a surface of a bead.

15 . The method of claim 12 , wherein the antibody is a soluble antibody.

16 . The method of claim 12 , wherein the antibody comprises at least one of an anti-CD3 antibody, an anti-CD28 antibody and an anti-CD2 antibody.

17 . The method of claim 1 , wherein the vector is a lentiviral vector or a retrovirus.

18 . The method of claim 1 , wherein the expanding comprises at least one or more of feeding, washing, monitoring, and selecting of the transduced immune cell culture.

19 . The method of claim 1 , wherein an oxygen level of the transduced immune cell culture is optimized for the immune cell culture.

20 . The method of claim 1 , wherein the cell engineering system recirculates cell culture media through an oxygenation component during one or more of steps (a) to (e).

21 . The method of claim 1 , wherein the cell engineering system recirculates nutrients, waste, released cytokines, and/or dissolved gasses during steps (a) to (e).

22 . The method of claim 1 , wherein a carbon dioxide level provided by the cell engineering system decreases during step (c).

23 . The method of claim 1 , wherein the cell engineering system is configured to perform several rounds of feeding, washing, monitoring, and selecting of the transduced immune cell culture.

24 . The method of claim 1 , wherein the concentrating comprises centrifugation, supernatant removal following sedimentation, or filtration.

25 . The method of claim 1 , wherein the cell engineering system comprises a plurality of chambers, and wherein each of steps (a) to (e) is performed in a different chamber of the plurality of chambers of the cell engineering system.

26 . The method of claim 1 , further comprising removing the activation reagent from the activated immune cell culture after step (a).

27 . The method of claim 1 , further comprising removing the vector following the transducing in (b).

28 . The method of claim 1 , wherein the cell engineering system contains the cell culture of (a), the activation reagent, the vector, and cell culture medium prior to starting the method.

29 . The method of claim 1 , wherein transduction efficiency in (b) of the method is at least 20% higher than the transduction efficiency of the method utilizing a flexible, gas permeable bag for cell culture.

30 . The method of claim 1 , wherein expansion of the transduced immune cell culture in (c) of the method produces at least 20% more genetically modified immune cells than a method utilizing manual cell culture with a flexible, gas permeable bag.

31 . The method of claim 1 , wherein the cell engineering system comprises a plurality of chambers, and wherein each of steps (a) to (e) is performed in a different chamber of the plurality of chambers of the cell engineering system, each of the activation reagent, the vector, and a cell culture medium are contained in a different chamber of the plurality of the chambers prior to starting the method, and wherein at least one of the plurality of chambers is maintained at a temperature for growing cells and at least one of the plurality of chambers is maintained at a refrigerated temperature.

Assignees

• Lonza Walkersville Inc
• Lonza Cologne Gmbh
• Octane Biotech Inc

Inventors

• Shi Yaling
• Mcafee Erika
• Bandapalle Samatha
• Siehoff Ann
• Gleissner Timo
• O'Connor Joseph
• Abraham Eytan
• Purpura Kelly
• Trainor Nuala
• Smith Timothy

Classifications

• C12N15/85

  for animal cells · CPC title

• C12M23/42

  Integrated assemblies, e.g. cassettes or cartridges · CPC title

• C12M41/00

  Means for regulation, monitoring, measurement or control, e.g. flow regulation (controlling or regulating chemical, physical or physicochemical processes B01J19/0006; heating or cooling apparatus for laboratory use B01L7/00; electro optical investigation of individual particles, flow cytometers G01N15/14; automatic analysis G01N35/00; controlling or regulating in general G06N) · CPC title

• C12N2510/00

  Genetically modified cells · CPC title

• C12M29/20

  Degassing; Venting; Bubble traps (means for collection or storage of gas C12M23/36; gas collection apparatus for laboratory use B01L5/02) · CPC title