Methods of Reducing Level of One or More Impurities in a Sample During Protein Purification

Patent metadata

Field	Value
Publication number	US-2016016992-A1
Application number	US-201514747029-A
Country	US
Kind code	A1
Filing date	Jun 23, 2015
Priority date	Aug 19, 2011
Publication date	Jan 21, 2016
Grant date	—

Abstract

The present invention provides novel and improved protein purification processes which incorporate certain types of carbonaceous materials and result in effective and selective removal of certain undesirable impurities without adversely effecting the yield of the desired protein product.

First claim

1 - 43 . (canceled)

44 . A method of reducing the level of one or more impurities in a sample containing an antibody of interest and one or more impurities, the method comprising the steps of:
(i) contacting a sample comprising an antibody and one or more impurities with a chromatography column containing an affinity media; (ii) obtaining a first eluate of the sample; (iii) contacting the first eluate from (ii) in flow-through mode with a carbonaceous material; (iv) obtaining a second eluate of the sample; (v) contacting the second eluate with an anion exchange porous media; and (vi) obtaining a third eluate of the sample,
wherein the third eluate comprises a lower level of one or more impurities relative to the level of one or more impurities when the carbonaceous material is not used in the method.

45 . A flow-through process for purifying an antibody from a Protein A eluate comprising the steps of:
(i) contacting the eluate recovered from a Protein A chromatography column with activated carbon; (ii) contacting the flow-through sample from step (i) with an anion exchange chromatography media; (iii) contacting the flow-through sample from step (ii) with a cation exchange chromatography media; and (iv) obtaining the flow-through sample from step (iii) comprising the antibody,
wherein the eluate flows continuously through steps (i)-(iii) and wherein
level of one or more impurities in the flow-through sample after step (iii) is lower than the level in the eluate in step (i).

46 . The flow-through process of claim 44 , further comprising subjecting the flow-through sample from step (vi) to virus filtration.

47 . The flow-through process of claim 45 , further comprising subjecting the flow-through sample from step (iv) to virus filtration.

48 . The flow-through process of claim 45 , further comprising use of an in-line static mixer and/or a surge tank between steps (ii) and (iii) to change pH.

49 . The flow-through process of claim 44 , wherein the process employs a single skid.

50 . The flow-through process of claim 45 , wherein the process employs a single skid.

51 . The flow-through process of claim 44 , wherein the eluate in step (ii) is subjected to virus inactivation prior to contacting with carbonaceous material.

52 . The flow-through process of claim 45 , wherein the eluate from the Protein A chromatography column is subjected to virus inactivation prior to contacting with activated carbon.

53 . The method of claim 44 , wherein the antibody is a monoclonal antibody or a polyclonal antibody.

54 . The method of claim 45 , wherein the antibody is a monoclonal antibody or a polyclonal antibody.

55 . The method of claim 44 , wherein carbonaceous material comprises activated carbon.

56 . The method of claim 55 , wherein the activated carbon comprises activated charcoal.

57 . The method of claim 45 , wherein the activated carbon comprises activated charcoal.

58 . The method of claim 45 , wherein the anion exchange chromatography media and/or the cation exchange chromatography media is packed in a column.

59 . The method of claim 45 , wherein the anion exchange chromatography media comprises an anion exchange resin.

60 . The method of claim 45 , wherein the cation exchange chromatography media comprises a cation exchange resin.

61 . The method of claim 44 , wherein the carbonaceous material is packed in a column, a sealed disposable device, a cartridge or a capsule.

62 . The method of claim 45 , wherein the activated carbon is packed in a column, a sealed disposable device, a cartridge or a capsule.

63 . The method of claim 44 , wherein the carbonaceous material is impregnated into a porous material.

64 . The method of claim 45 , wherein the activated carbon is impregnated into a porous material.

65 . The method of claim 63 , wherein the porous material is contained within a column, a sealed disposable device, a cartridge or a capsule.

66 . The method of claim 64 , wherein the porous material is contained within a column, a sealed disposable device, a cartridge or a capsule.

67 . The method of claim 45 , wherein the anion exchange chromatography media is a porous adsorptive media having a surface coating comprising one or more polymeric primary amines or copolymers thereof.

68 . The method of claim 45 , wherein the anion exchange chromatography media is a porous adsorptive media having a surface coating comprising one or more primary, secondary, tertiary and quaternary amines.

69 . The method of claim 44 , wherein the anion exchange porous media is a membrane adsorber.

70 . A method of reducing the level of one or more impurities in a sample containing an antibody, the method comprising the steps of:
(i) providing a sample comprising an antibody and one or more impurities; (ii) contacting the sample with a suitable media to capture the antibody; (iii) obtaining a first eluate of the sample; (iv) contacting the first eluate with a carbonaceous material; (v) obtaining a second eluate of the sample; and (vi) contacting the second eluate with an anion exchange chromatography media; and (vii) obtaining a third eluate of the sample,
wherein the third eluate comprises a lower level of one or more impurities relative to the level of one or more impurities when step (iv) is not performed.

71 . The method of claim 70 , wherein the antibody is a monoclonal antibody or a polyclonal antibody.

72 . The method of claim 70 , wherein the anion exchange chromatography media is a resin or a membrane.

73 . The method of claim 70 , wherein the carbonaceous material is activated carbon.

Assignees

• Emd Millipore Corp

Inventors

• Bian Nanying
• Gillespie Christopher
• Stone Matthew T
• Kozlov Mikhail
• Chen Jie
• Siwak Martin

Classifications

• B01D15/363

  Anion-exchange · CPC title

• B01D15/1871

  placed in series · CPC title

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• B01J41/20

  Anion exchangers for chromatographic processes · CPC title

• C07K2317/10

  characterized by their source of isolation or production · CPC title