公开(公告)号：US20240327453A1

公开(公告)日：2024-10-03

申请号：US18442665

申请日：2024-02-15

Applicant: AMGEN INC.

Inventor： Ashish Sharma ,  Balakumar Thangaraj

IPC: C07K1/20 ,  C07K14/715

CPC classification number: C07K1/20 ,  C07K14/7151 ,  C07K2319/30

Abstract: The invention pertains to methods of purifying fusion proteins, in particular TNFR:Fc fusion proteins. Methods disclosed herein can be used to produce highly pure TNFR:Fc fusion proteins (e.g., etanercept) having a biological activity by removing hard to separate product related impurities such as clipped and/or mis-fold/aggregated TNFR:Fc fusion proteins.

公开(公告)号：US11952399B2

公开(公告)日：2024-04-09

申请号：US16708987

申请日：2019-12-10

Applicant: AMGEN INC.

Inventor： Ashish Sharma ,  Balakumar Thangaraj

IPC: C07K1/20 ,  C07K14/715

CPC classification number: C07K1/20 ,  C07K14/7151 ,  C07K2319/30

Abstract: The invention pertains to methods of purifying fusion proteins, in particular TNFR:Fc fusion proteins. Methods disclosed herein can be used to produce highly pure TNFR:Fc fusion proteins (e.g., etanercept) having a biological activity by removing hard to separate product related impurities such as clipped and/or mis-fold/aggregated TNFR:Fc fusion proteins.

公开(公告)号：US20220396599A1

公开(公告)日：2022-12-15

申请号：US17775386

申请日：2020-11-13

Applicant: AMGEN INC.

Inventor： Yan Wang ,  Nicholas Marchand ,  Robert Perry Brake ,  Maria Perry ,  Ashish Sharma ,  Eugene Sun

IPC: C07K1/34 ,  C07K16/28

Abstract: The present disclosure provides a downstream manufacturing method for the production of bispecific antigen-binding molecule products, which comprise at least two different binding domains. The process comprises at least one filtration step, wherein the filtration is ultrafiltration/diafiltration (UF/DF), and/or viral filtration (VF), wherein a β-cyclodextrin is applied in the process either in a buffer applied in the filtration step or added to the filtration pool after the UF/DF filtration step wherein the buffer or the pool have a concentration of about 0.1 to 3% (m/v) of a (3-cyclodextrin, to preferably keep the product in a non-aggregated form.

公开(公告)号：US20220306741A1

公开(公告)日：2022-09-29

申请号：US17641736

申请日：2020-09-10

Applicant: AMGEN INC.

Inventor： Ashish Sharma ,  Balakumar Thangaraj

IPC: C07K16/28 ,  C07K1/14

Abstract: The present invention provides a downstream purification method process for the production of bispecific antigen-binding polypeptides. The method comprises at least the steps of (i) providing a separation resin comprising a polymer matrix part and a ligand part, wherein the matrix part comprises polymethacrylate and has a particle size of about 30 to 60 pm, wherein the ligand part comprises recombinant protein L, and wherein the ligand part's protein L is covalently bound to the matrix part's particles, (ii) contacting a process fluid comprising the bispecific antigen-binding polypeptide with the separation resin, (iii) capturing the bispecific antigen-binding polypeptide by the ligand part of the separation resin, wherein the bispecific antigen-binding polypeptide reversibly binds to the ligand part of the separation resin, and wherein the remainder of the process fluid does not bind to the ligand part of the separation resin, (iv) washing the bound bispecific antigen-binding polypeptide with a wash buffer which does not elute the bispecific antigen-binding polypeptide from the ligand portion, and (v) elute the bispecific antigen-binding polypeptide from the ligand part with an elution buffer at a low pH.