B-cell cultivation method

Patent metadata

Field	Value
Publication number	US-10889802-B2
Application number	US-201816144240-A
Country	US
Kind code	B2
Filing date	Sep 27, 2018
Priority date	Mar 30, 2016
Publication date	Jan 12, 2021
Grant date	Jan 12, 2021

Abstract

Herein is reported a method for co-cultivating one or more B-cells comprising the step of incubating the one or more B-cells with EL4-B5 cells, whereby the EL4-B5 cells have been obtained/are from a cultivation of EL4-B5 cells that has a cell density of from 600,000 cells/ml up to 1,500,000 cells/ml.

First claim

The invention claimed is:

1. A method for co-cultivating one or more B-cells comprising the steps of
cultivating EL4-B5 cells to a cell density of more than 1,000,000 cells/ml to 1,500,000 cells/ml, and
incubating the one or more B-cells with an aliquot of the EL4-B5 cells obtained in the previous step.

2. The method according to claim 1 , wherein the cultivation of the EL4-B5 cells is to a cell density of 650,000 cells/ml to 1,450,000 cells/ml.

3. The method according to claim 1 , wherein the cultivation of the EL4-B5 cells is to a cell density of 1,400,000 cells/ml to 1,500,000 cells/ml.

4. The method according to claim 1 , wherein the incubating is additionally in the presence of a feeder mix.

5. The method according to claim 4 , wherein the feeder mix comprises one or more of:
i) interleukin-1 beta and tumor necrosis factor alpha,
ii) interleukin-2 (IL-2) and/or interleukin-10 (IL-10),
iii) Staphylococcus aureus strain Cowan's cells (SAC),
iv) interleukin-21 (IL-21) and optionally interleukin-2 (IL-2),
v) B-cell activation factor of the tumor necrosis factor family (BAFF),
vi) interleukin-6 (IL-6),
vii) interleukin-4 (IL-4), and
viii) thymocyte cultivation supernatant.

6. The method according to claim 4 , wherein the feeder mix comprises IL-1ß, TNF-α, IL-10 and one or more of the members selected from the group consisting of: IL-21, SAC, BAFF, IL-2, IL-4, and IL-6.

7. The method according to claim 1 , wherein the EL4-B5 cells are, after the cultivating and prior to the incubating, irradiated with y-radiation.

8. The method according to claim 1 , wherein the method is for the co-cultivation of one B-cell.

9. The method according to claim 8 , wherein the one B-cell is a single deposited B-cell.

10. The method according to claim 1 , wherein the incubating is for 5 to 14 days.

11. A method for producing an antibody, the method comprising performing the method according to claim 1 , thereby producing the antibody.

12. The method according to claim 11 , wherein the antibody is produced by the one or more B cells.

13. The method according to claim 12 , wherein the antibody is produced by the one or more B cells and secreted into cell culture supernatant.

14. The method according to claim 12 , further comprising recovering the antibody from the one or more B cells.

15. The method according to claim 13 , further comprising recovering the antibody from the cell culture supernatant.

Assignees

• Hoffmann La Roche

Inventors

• Offner Sonja
• Jung Friederike

Classifications

• C12N5/0635 Primary

  B lymphocytes · CPC title

• C12N2502/1185

  Thymus cells · CPC title

• C12N2525/00

  Culture process characterised by gravity, e.g. microgravity · CPC title

• C12N2510/02

  Cells for production · CPC title

• C12N2502/30

  tumour cells · CPC title