公开(公告)号：US10995113B2

公开(公告)日：2021-05-04

申请号：US16893574

申请日：2020-06-05

Applicant: Cytiva BioProcess R&D AB

Inventor： Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: A23J1/00 ,  C07K1/22 ,  B01D15/38 ,  B01J20/24 ,  B01J20/28 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/06 ,  C07K17/10

Abstract: The invention relates to a separation matrix comprising at least 11 mg/ml Fc-binding ligands covalently coupled to a porous support, wherein: a) the ligands comprise multimers of alkali-stabilized Protein A domains, and b) the porous support comprises cross-linked polymer particles having a volume-weighted median diameter (d50,v) of 56-70 micrometers and a dry solids weight of 55-80 mg/ml.

公开(公告)号：US12134633B2

公开(公告)日：2024-11-05

申请号：US18298857

申请日：2023-04-11

Applicant: Cytiva BioProcess R&D AB

Inventor： Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: C07K1/22 ,  B01D15/38 ,  B01J20/24 ,  B01J20/28 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/06 ,  C07K17/10

Abstract: The invention relates to a separation matrix comprising at least 11 mg/ml Fc-binding ligands covalently coupled to a porous support, wherein: a) the ligands comprise multimers of alkali-stabilized Protein A domains, and b) the porous support comprises cross-linked polymer particles having a volume-weighted median diameter (d50,v) of 56-70 micrometers and a dry solids weight of 55-80 mg/ml.

公开(公告)号：US20240018184A1

公开(公告)日：2024-01-18

申请号：US18354413

申请日：2023-07-18

Applicant: CYTIVA BIOPROCESS R&D AB

Inventor： Annika Forss ,  Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: C07K1/22 ,  B01J20/26 ,  B01J20/285 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/12 ,  C07K17/10

CPC classification number: C07K1/22 ,  B01J20/267 ,  B01J20/285 ,  B01J20/286 ,  B01J2220/52 ,  C07K14/31 ,  C07K16/00 ,  C07K16/1271 ,  C07K17/10 ,  B01J20/3274

Abstract: The present invention concerns a method of cleaning and/or sanitizing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support. The method comprises the steps of:

a) optionally purifying a mixture comprising a first immunoglobulin using the separation matrix;
b) providing a cleaning liquid comprising at least 50% by volume of an aqueous alkali metal hydroxide solution; and
c) cleaning and/or sanitizing the separation matrix by contacting the cleaning liquid with the separation matrix for a predetermined contact time.


The alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO 51 or SEQ ID NO 52, wherein the amino acid residues at positions 13 and 44 of SEQ ID NO 51 or 52 are asparagines and wherein at least the asparagine residue at position 3 of SEQ ID NO 51 or 52 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.

公开(公告)号：US11708390B2

公开(公告)日：2023-07-25

申请号：US16095721

申请日：2017-05-10

Applicant: Cytiva BioProcess R&D AB

Inventor： Annika Forss ,  Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: C07K1/22 ,  B01D15/00 ,  C07K14/31 ,  B01J20/286 ,  B01J20/32 ,  C07K16/06 ,  C07K16/12 ,  C07K17/10

CPC classification number: C07K1/22 ,  B01D15/00 ,  B01J20/286 ,  B01J20/3212 ,  B01J20/3274 ,  C07K14/31 ,  C07K16/065 ,  C07K16/1271 ,  C07K17/10 ,  B01J2220/52

Abstract: The present invention concerns a method of storing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support. The method comprises the steps of: a) providing a storage liquid comprising at least 50% by volume of an aqueous alkali metal hydroxide solution; b) permeating the separation matrix with the storage liquid; and c) storing the storage liquid-permeated separation matrix for a storage time of at least days. The alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA), as defined by, or having at least 80% such as at least 90%, 95% or 98% identity to, SEQ ID NO 51 or SEQ ID NO 52, wherein the amino acid residues at positions 13 and 44 of SEQ ID NO 51 or 52 are asparagines and wherein at least the asparagine residue at position 3 of SEQ ID NO 51 or 52 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.

公开(公告)号：US20230257417A1

公开(公告)日：2023-08-17

申请号：US18181974

申请日：2023-03-10

Applicant: Cytiva BioProcess R&D AB

Inventor： Annika Kristina Forss ,  Gustav Jose Rodrigo ,  Tomas Bjorkman ,  Jesper Ulf Hansson ,  Mats Ander

IPC: C07K1/22 ,  C07K16/12 ,  C07K17/10 ,  B01J20/286 ,  C07K14/31 ,  C07K16/06 ,  B01D15/00 ,  B01J20/32

CPC classification number: C07K1/22 ,  C07K16/1271 ,  C07K17/10 ,  B01J20/286 ,  C07K14/31 ,  C07K16/065 ,  B01D15/00 ,  B01J20/3274 ,  B01J20/3212 ,  B01J2220/52

Abstract: The invention relates to a method of isolating an immunoglobulin, comprising the steps of: a) providing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support: b) contacting a liquid sample comprising an immunoglobulin with the separation matrix; c) washing said separation matrix with a washing liquid; d) eluting the immunoglobulin from the separation matrix with an elution liquid, and e) cleaning the separation matrix with a cleaning liquid, wherein the alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA).

公开(公告)号：US11753438B2

公开(公告)日：2023-09-12

申请号：US16095753

申请日：2017-05-10

Applicant: CYTIVA BIOPROCESS R&D AB

Inventor： Annika Forss ,  Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: C07K1/22 ,  B01J20/26 ,  B01J20/285 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/12 ,  C07K17/10

CPC classification number: C07K1/22 ,  B01J20/267 ,  B01J20/285 ,  B01J20/286 ,  B01J20/3274 ,  C07K14/31 ,  C07K16/00 ,  C07K16/1271 ,  C07K17/10 ,  B01J2220/52

Abstract: The present invention concerns a method of cleaning and/or sanitizing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support. The method comprises the steps of: a) optionally purifying a mixture comprising a first immunoglobulin using the separation matrix; b) providing a cleaning liquid comprising at least 50% by volume of an aqueous alkali metal hydroxide solution; and c) cleaning and/or sanitizing the separation matrix by contacting the cleaning liquid with the separation matrix for a predetermined contact time. The alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO 51 or SEQ ID NO 52, wherein the amino acid residues at positions 13 and 44 of SEQ ID NO 51 or 52 are asparagines and wherein at least the asparagine residue at position 3 of SEQ ID NO 51 or 52 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.

公开(公告)号：US20230279047A1

公开(公告)日：2023-09-07

申请号：US18298857

申请日：2023-04-11

Applicant: Cytiva BioProcess R&D AB

Inventor： Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: C07K1/22 ,  B01D15/38 ,  B01J20/24 ,  B01J20/28 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/06 ,  C07K17/10

CPC classification number: C07K1/22 ,  B01D15/3809 ,  B01J20/24 ,  B01J20/28016 ,  B01J20/28078 ,  B01J20/286 ,  B01J20/3212 ,  B01J20/3274 ,  C07K14/31 ,  C07K16/00 ,  C07K16/065 ,  C07K17/10

Abstract: The invention relates to a separation matrix comprising at least 11 mg/ml Fc-binding ligands covalently coupled to a porous support, wherein:

a) the ligands comprise multimers of alkali-stabilized Protein A domains, and
b) the porous support comprises cross-linked polymer particles having a volume-weighted median diameter (d50,v) of 56-70 micrometers and a dry solids weight of 55-80 mg/ml.

公开(公告)号：US20210187476A1

公开(公告)日：2021-06-24

申请号：US17252378

申请日：2019-06-25

Applicant: Cytiva BioProcess R&D AB

Inventor： Johan Fredrik Öhman ,  Jesper Ulf Hansson ,  Jasmin Faroque ,  Eva Holmgren ,  David Bror Lennart Jansson

IPC: B01J20/24 ,  B01J20/28 ,  B01J20/288 ,  B01J20/30 ,  B01J20/32 ,  C08B37/00 ,  B01J20/281

Abstract: The present invention relates to chromatography beads, production and use thereof. More closely the invention relates to small, rigid and nan-permeable agarose beads suitable for example as stationary phase in high performance liquid chromatography (HPLC) for analyses of biomolecules, such as, peptides and proteins; and methods for producing such beads.

公开(公告)号：US11685764B2

公开(公告)日：2023-06-27

申请号：US17221438

申请日：2021-04-02

Applicant: Cytiva BioProcess R&D AB

Inventor： Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander ,  Jesper Ulf Hansson

IPC: A23J1/00 ,  C07K1/22 ,  B01D15/38 ,  B01J20/24 ,  B01J20/28 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/06 ,  C07K17/10

CPC classification number: C07K1/22 ,  B01D15/3809 ,  B01J20/24 ,  B01J20/286 ,  B01J20/28016 ,  B01J20/28078 ,  B01J20/3212 ,  B01J20/3274 ,  C07K14/31 ,  C07K16/00 ,  C07K16/065 ,  C07K17/10

Abstract: The invention relates to a separation matrix comprising at least 11 mg/ml Fc-binding ligands covalently coupled to a porous support, wherein:



a) the ligands comprise multimers of alkali-stabilized Protein A domains, and
b) the porous support comprises cross-linked polymer particles having a volume-weighted median diameter (d50, v) of 56-70 micrometers and a dry solids weight of 55-80 mg/ml.

公开(公告)号：US11623941B2

公开(公告)日：2023-04-11

申请号：US16884475

申请日：2020-05-27

Applicant: Cytiva BioProcess R&D AB

Inventor： Annika Kristina Forss ,  Gustav Jose Rodrigo ,  Tomas Bjorkman ,  Jesper Ulf Hansson ,  Mats Ander

IPC: C07K1/22 ,  B01D15/08 ,  C07K16/12 ,  C07K17/10 ,  B01J20/286 ,  C07K14/31 ,  C07K16/06 ,  B01D15/00 ,  B01J20/32

Abstract: The invention relates to a method of isolating an immunoglobulin, comprising the steps of: a) providing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support, b) contacting a liquid sample comprising an immunoglobulin with the separation matrix, c) washing said separation matrix with a washing liquid, d) eluting the immunoglobulin from the separation matrix with an elution liquid, and e) cleaning the separation matrix with a cleaning liquid, wherein the alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO: 51 or SEQ ID NO: 52, wherein the amino acid residues at positions 13 and 44 of SEQ ID NO: 51 or 52 are asparagines and wherein at least the asparagine residue at position 3 of SEQ ID NO: 51 or 52 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.