Processing

Please wait...

Settings

Settings

Goto Application

1. WO2020141145 - ANTI-RABBIT CD19 ANTIBODIES AND METHODS OF USE

Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

[ EN ]

Patent Claims

1. An antibody binding to rabbit CD 19 comprising

(a) a HVR-H1 comprising the amino acid sequence of SEQ ID NO: 32 or 33 or 34,

(b) a HVR-H2 comprising the amino acid sequence of SEQ ID NO: 35 or 36,

(c) a HVR-H3 comprising the amino acid sequence of SEQ ID NO: 37,

(d) a HVR-L1 comprising the amino acid sequence of SEQ ID NO: 38,

(e) a HVR-L2 comprising the amino acid sequence of SEQ ID NO: 39, and

(f) a HVR-L3 comprising the amino acid sequence of SEQ ID NO: 40.

2. The antibody according to claim 1, wherein the antibody is a monoclonal antibody.

3. The antibody according to any one of claims 1 to 2, wherein the antibody is a chimeric or humanized antibody.

4. The antibody according to any one of claims 1 to 2, wherein the antibody comprises a variable heavy chain domain of SEQ ID NO: 30 and a variable light chain domain of SEQ ID NO: 26.

5. The antibody according to any one of claims 1 to 4, wherein the antibody is a full length antibody or an antibody fragment.

6. The antibody according to any one of claims 1 to 5, wherein the antibody is conjugated to a detectable label.

7. The antibody according to claim 6, wherein the detectable label is a fluorescent dye.

8. A method for selecting a rabbit B-cell comprising the following steps:

a) incubating a multitude of rabbit B-cells with an antibody according to any one of claims 1 to 7,

b) selecting one or more B-cells to which the antibody according to any one of claims 1 to 7 is bound and thereby selecting a rabbit B-cell.

9. The method according to claim 8 further comprising one or more of the following steps:

after step b) and prior to step c): incubating the rabbit B-cells at 37 °C for one hour in co-cultivation medium, and/or

c) single depositing one or more rabbit B-cells to which the antibody according to any one of claims 1 to 7 is bound, and/or

d) co-cultivating the single deposited rabbit B-cells with feeder cells in a co-cultivation medium, and/or

e) selecting a rabbit B-cell proliferating in step d) and thereby selecting a rabbit B -cell.

10 A method for removing non B-cells for a cultivation comprising the following steps:

a) co-cultivating rabbit B-cells, which have been deposited either as single cells or as pool of cells, with feeder cells,

b) incubating the cells from the co-cultivation obtained in step a) with the antibody according to any one of claims 1 to 7, and

c) selecting one or more rabbit B-cells to which the antibody according to any one of claims 1 to 7 is bound and thereby removing non-B-cells.

11. A method for determining the number B-cells in a co-cultivation of a single deposited B-cell with feeder cells comprising the following steps:

a) co-cultivating a single deposited rabbit B-cell with feeder cells,

b) incubating the cells from the co-cultivation obtained in step a) with the antibody according to any one of claims 1 to 7, and

c) determining the number of B-cells in the cultivation by counting the number of cells to which the antibody according to any one of claims 1 to 7 is bound.

12. A method for co-cultivating one or more rabbit B-cells comprising the steps of

incubating a multitude of rabbit B-cells/labelling individual B-cells of a multitude of rabbit B-cells with an antibody according to any one of claims 1 to 7,

selecting/depositing one or more rabbit B-cells that have the antibody according to any one of claims 1 to 7 bound to their surface/that have been labelled either as individual B-cells (single deposited B-cell) or as a pool of B-cells, and

- co-cultivating the single deposited rabbit B-cells or the pool of rabbit B- cells with feeder cells.

13. The method according to any one of claims 9 to 12, wherein the co-cultivating is in the presence of a synthetic feeder mix that comprises IL-Ib, TNFa, IL- 10, and one or more selected from IL-21, SAC, BAFF, IL-2, IL-4, and IL-6.

14. A method for selecting B-cells comprising the following steps:

a) obtaining B-cells from the blood of a rabbit,

b) incubating the B-cells with an antibody according to any one of claims 1 to 5, which is bound to a bead,

c) removing non-bound B-cells,

d) recovering the bound B-cells from the beads and thereby selecting one or more B-cells.

15. The method according to claim 14 further comprising the following steps

e) optionally incubating the recovered B-cells at 37 °C for one hour in co cultivation medium,

f) depositing one or more recovered B-cells in an individual container,

g) co-cultivating the deposited cells with a feeder cell in a co-cultivation medium,

h) selecting a B-cell proliferating in step g) and thereby selecting a B-cell.