Methods of reducing the enzymatic hydrolysis activity rate in a composition obtained from a purification platform

Patent metadata

Field	Value
Publication number	US-12479883-B2
Application number	US-202117517531-A
Country	US
Kind code	B2
Filing date	Nov 2, 2021
Priority date	May 3, 2019
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

The present disclosure provides purification platforms comprising a depth filter step and/or a hydrophobic interaction chromatography (HIC) step. Also disclosed herein are methods of using the purification platforms described herein and compositions obtained therefrom, such as pharmaceutical compositions.

First claim

What is claimed is:

1 . A method of reducing an enzymatic hydrolysis activity rate of a composition obtained from a purification platform, the method comprising subjecting a sample to the purification platform comprising, in order:
(a) a capture step comprising processing via affinity chromatography; and (b) a purification step comprising processing via a chromatography selected from the group consisting of a HIC, a cation exchange chromatography, and a multimodal chromatography, wherein the purification platform further comprises one or more depth filtration steps,
wherein the one or more depth filtration steps are performed at any one or more of: prior to the capture step; after the capture step; or after the capture step and prior to the purification step,
wherein each depth filtration step comprises processing via a depth filter, and
wherein the depth filter comprises materials selected from the group consisting of:
(i) a silica filter aid and a polyacrylic fiber pulp; and
(ii) cellulose fibers, diatomaceous earth, and perlite,
wherein the depth filter comprising the cellulose fibers, diatomaceous earth, and perlite comprises two layers, wherein each layer comprises a cellular filter matrix impregnated with a filter aid comprising one or more of diatomaceous earth or perlite, and wherein each layer further comprises a resin binder,

thereby reducing the enzymatic hydrolysis activity rate of the composition as compared to purification of the sample using the same purification platform without the one or more depth filtration steps.

2 . The method of claim 1 , wherein the relative reduction of the enzymatic hydrolysis activity rate of the composition, as compared to purification of the sample using the same purification platform without the depth filtration step, is at least about 20%.

3 . The method of claim 1 , wherein the level of one or more hydrolytic enzymes in the composition obtained from the purification platform is reduced as compared to purification of the sample using the same purification platform without the one or more depth filtration steps, and wherein the relative reduction of the level of one or more hydrolytic enzymes in the composition, as compared to purification of the sample using the same purification platform without the depth filtration step, is at least about 20%.

4 . The method of claim 1 , wherein the degradation of a polysorbate in the composition obtained from the purification platform is reduced as compared to purification of the sample using the same purification platform without the one or more depth filtration steps, and wherein the relative reduction of degradation of the polysorbate in the composition, as compared to purification of the sample using the same purification platform without the depth filtration step, is at least about 5%.

5 . The method of claim 1 , wherein the depth filter is selected based on the pH of the solution entering the depth filter, and wherein the depth filter comprising the silica filter aid and the polyacrylic fiber pulp is selected when the solution entering the depth filter is about 5 to about 6.5.

6 . The method of claim 1 , wherein the HIC is a phenyl SEPHAROSE® fast flow chromatography.

7 . The method of claim 1 , wherein the cation exchange chromatography is POROS®50HS.

8 . The method of claim 1 , wherein the purification platform further comprises a second depth filtration step comprising processing via the depth filter comprising the silica filter aid and the polyacrylic fiber pulp, and wherein the second depth filtration step occurs after the capture step and prior to the purification step.

9 . The method of claim 1 , wherein the multimodal chromatography is Capto Adhere.

10 . The method of claim 1 , wherein the purification platform further comprises a second depth filtration step comprising processing via a depth filter comprising a hydrogel Q-functionalized non-woven media and a multi-zone microporous membrane, and wherein the second depth filtration step occurs after the capture step and prior to the purification step.

11 . The method of claim 1 , wherein the purification platform is for purification of a target from the sample, wherein the sample comprises the target and one or more host cell impurities, and wherein the host cell impurity is a host cell protein.

12 . The method of claim 1 , wherein the purification platform further comprises a virus inactivation step, wherein the virus inactivation step is performed after the capture step.

13 . The method of claim 12 , wherein the one or more depth filtration steps are performed after the virus inactivation step.

14 . The method of claim 1 , wherein the purification platform further comprises an ultrafiltration/diafiltration (UFDF) step, and wherein the UFDF step is performed after the purification step.

15 . The method of claim 1 , further comprising determining the enzymatic hydrolysis activity rate of the composition.

16 . The method of claim 1 , further comprising determining the level of one or more hydrolytic enzymes in the composition.

17 . The method of claim 1 , wherein the sample comprises one or more host cell proteins, wherein one of the one or more host cell proteins is a hydrolytic enzyme, and wherein the hydrolytic enzyme is a lipase, an esterase, a thioesterase, a phospholipase, or a ceramidase.

18 . The method of claim 1 , wherein the affinity chromatography is a protein A chromatography from FcXL affinity chromatography or MABSELECT SURE™.

Assignees

• Genentech Inc
• Hoffmann La Roche

Inventors

• Leiss Michael
• Meyer Christian
• Morris Alexandra Thomas
• Spensberger Bernhard
• Yigzaw Yinges
• Edelmann Franziska
• Falkenstein Roberto
• Graf Tobias
• Gu Jie
• Prabhakar Jeevan

Classifications

• C07K1/36

  by a combination of two or more processes of different types · CPC title

• C07K1/34

  by filtration, ultrafiltration or reverse osmosis · CPC title

• C07K1/20 Primary

  Partition-, reverse-phase or hydrophobic interaction chromatography · CPC title

• C07K1/165

  mixed-mode chromatography · CPC title

• B01D2315/16

  Diafiltration · CPC title