Novel antibody purification method, antibody obtained therefrom, novel antibody purification method using cation exchanger, and antibody obtained therefrom

2:

• A method for purifying an antibody or a substance derived from an antibody with a carrier 1 having an affinity ligand with affinity for the antibody or the substance derived from the antibody and a carrier 2 having a cation exchange group, wherein
• a solution containing the antibody or the substance derived from the antibody is applied to an integrated column 1 connecting a column containing the carrier 1 and a column containing the carrier 2 in series or a column 2 having a mixture of the carrier 1 and the carrier 2, and
• an eluate is passed through the integrated column or the mixed column to elute the adsorbed antibody or the substance derived from the antibody.

3:

• The method according to claim 1, wherein the antibody or the substance derived from the antibody is eluted by the eluate having an acidic pH with an ionic strength linear gradient.

4:

• The method according to claim 1, wherein the antibody or the substance derived from the antibody is eluted by the eluate having an acidic pH with an ionic strength step gradient.

7:

• The method according to claim 1, wherein the carrier 1 has protein A, protein G, protein L or an analog thereof as a ligand.

8:

• The method according to claim 1, wherein the carrier 1 has protein A or an analog thereof as a ligand.

9:

• The method according to claim 1, wherein the antibody or the substance derived from the antibody is immunoglobulin G, immunoglobulin G derivative, or Fc-containing molecule.

10:

• The method according to claim 1, wherein the antibody or the substance derived from the antibody is Fab, scFv, diabody, or a molecule containing a binding part to an antigen.

11:

• The method according to claim 1, wherein the carrier 2 has a carboxyl group as a ligand.

12:

• The method according to claim 11, wherein the carboxyl group is derived from an acidic amino acid.

13:

• The method according to claim 1, wherein the elution pH of the antibody or the substance derived from the antibody is less than 5.0.

14:

• The method according to claim 1, wherein 10% DBC of the carrier 1 to IgG at 6 minutes of contact time is 1 mg/mL or more and 100 mg/mL or less.

15:

• The method according to claim 1, wherein the carrier 2 has the ion exchange capacity of 0.001 mmol/mL or more and 0.5 mmol/mL or less.

16:

• The method according to claim 1, wherein the carrier 1 has the volume average particle diameter of 1 μm or more and 1000 μm or less, and the carrier 2 has the volume average particle diameter of 1 μm or more and 1000 μm or less.

17:

• The method according to claim 1, wherein
• the solution containing the antibody or the substance derived from the antibody is applied to the carrier 2 having a cation exchange group having a carboxyl group-containing ligand and pKa of 4.0 or more, and
• the antibody or the substance derived from the antibody is eluted by an acidic buffer having pH of 4.0 or less.

18:

• The method according to claim 1, wherein 10% DBC of the carrier 2 to IgG at 6 minutes of contact time is 1 mg/mL or more and 200 mg/mL or less.

20:

• The method according to claim 2, wherein the ratio of the carrier 1 to the carrier 2 of the integrated column is 1/20 or more and 20/1 or less on the basis of the volume.

22:

• The method according to claim 2, wherein the ratio of the carrier 1 to the carrier 2 of the mixed column is 1/20 or more and 20/1 or less on the basis of the volume.

23:

• The method according to claim 1, wherein the ratio of 10% DBC of the carrier 2 to IgG at 6 minutes of contact time relative to 10% DBC of the carrier 1 to IgG at 6 minutes of contact time under each adsorption condition is 1/10 times or more and 10 times or less.

24:

• An antibody or a substance derived from an antibody purified with the method according to claim 1.

25:

• A method for using a carrier having a cation exchange group, wherein
• a solution containing an antibody or a substance derived from an antibody is applied to a carrier 2 having a cation exchange group having a carboxyl group-containing ligand and pKa of 4.0 or more, and
• the antibody or the substance derived from the antibody is eluted by an acidic buffer having pH of 4.0 or less.

26:

• The method according to claim 25, wherein the carboxyl group-containing ligand is derived from an acidic amino acid.

27:

• The method according to claim 25, wherein 10% DBC of the carrier 2 to IgG at 6 minutes of contact time is 1 mg/mL or more and 200 mg/mL or less.

28:

• The method according to claim 25, wherein the carrier 2 has the ion exchange capacity of 0.001 mmol/mL or more and 0.5 mmol/mL or less.

29:

• The method according to claim 25, wherein the carrier 2 has the volume average particle diameter of 1 μm or more and 1000 μm or less.

34:

• The method according to claim 25, wherein the antibody or the substance derived from the antibody is immunoglobulin G, immunoglobulin G derivative, Fc-containing molecule, Fab, scFv, diabody, or a molecule containing a binding part to an antigen.

35:

• The method according to claim 25, wherein the antibody or the substance derived from the antibody is eluted by the eluate having an acidic pH with an ionic strength linear gradient.

36:

• The method according to claim 25, wherein the antibody or the substance derived from the antibody is eluted by the eluate having an acidic pH with an ionic strength step gradient.

37:

• An antibody or a substance derived from an antibody purified with the method according to claim 25.