公开(公告)号：US12037359B2

公开(公告)日：2024-07-16

申请号：US17114773

申请日：2020-12-08

Applicant: CYTIVA BIOPROCESS R&D AB

Inventor： Gustav José Rodrigo ,  Tomas Bjorkman ,  Mats Ander

IPC: C07K1/22 ,  B01D15/38 ,  B01J20/24 ,  B01J20/26 ,  B01J20/28 ,  B01J20/285 ,  B01J20/286 ,  B01J20/32 ,  C07K14/31 ,  C07K16/00 ,  C07K16/06 ,  C07K16/12 ,  C07K17/10

CPC classification number: C07K1/22 ,  B01D15/3809 ,  B01J20/24 ,  B01J20/267 ,  B01J20/28016 ,  B01J20/28078 ,  B01J20/285 ,  B01J20/286 ,  B01J20/3212 ,  B01J20/3274 ,  C07K14/31 ,  C07K16/00 ,  C07K16/065 ,  C07K16/1271 ,  C07K17/10 ,  B01J2220/52

Abstract: An Fc-binding polypeptide of improved alkali stability, comprising a mutant of a parental Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 22, SEQ ID NO: 51 or SEQ ID NO: 52, wherein at least the asparagine or serine residue at the position corresponding to position 11 in SEQ ID NO: 4-7 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：US11136359B2

公开(公告)日：2021-10-05

申请号：US15900419

申请日：2018-02-20

Applicant: Cytiva BioProcess R&D AB

Inventor： Gustav Rodrigo ,  Mats Ander ,  Tomas Bjorkman ,  Goran Bauren

IPC: C07K14/31 ,  B01D15/38 ,  B01J20/286 ,  B01J20/32 ,  C07K1/22 ,  C07K14/745 ,  C07K16/00

Abstract: The invention discloses a polypeptide with improved alkaline stability, which polypeptide comprises a mutant of a B or C domain of Staphylococcus Protein A (SpA), as specified by SEQ ID NO 1 or SEQ ID NO 2, or of Protein Z, as specified by SEQ ID NO 3, wherein at least the glutamine residue at position 9 has been mutated to an amino acid other than asparagine. The invention also discloses multimers of said polypeptide, as well as separation matrices comprising the multimers or polypeptides.

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：US20240391984A1

公开(公告)日：2024-11-28

申请号：US18682409

申请日：2022-08-24

Applicant: Cytiva Bioprocess R&D AB

Inventor： Mats Ander ,  Joakim Galli ,  Andreas L. Jonsson ,  Gustav Rodrigo

IPC: C07K16/12 ,  C07K1/22

Abstract: The present relates to a polypeptide that binds to an immunoglobulin or a fragment thereof. More specifically, it relates to a kappa light-chain binding polypeptide with high binding affinity and improved alkali stability. The one kappa light-chain binding comprises a mutated binding domain of Peptostreptococcus Protein L, derived from any one of the amino acid sequences SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17 or SEQ ID NO:18, said amino acid sequences having N6H, N41H and N56Y or N56Q mutations.

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：US20240228530A1

公开(公告)日：2024-07-11

申请号：US18562092

申请日：2022-06-22

Applicant: Cytiva BioProcess R&D AB

Inventor： Bengt Westerlund ,  Mats Ander

IPC: C07K1/22

CPC classification number: C07K1/22 ,  C07K2317/31 ,  C07K2317/56

Abstract: The present invention relates to a method of separating bispecific antibodies or bispecific antibody fragments. The method comprises the steps of a) providing a feed comprising bispecific antibodies or bispecific antibody fragments; b) contacting the feed with a separation matrix having affinity ligands coupled to a support; c) optionally washing the separation resin with a washing liquid; d) applying an elution buffer to the separation resin, to elute the antibodies or antibody fragments bound to the affinity ligand; wherein in step d) a pH gradient is applied over the elution buffer, said pH gradient being from about 6 to about 2.

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：US20220315647A1

公开(公告)日：2022-10-06

申请号：US17604964

申请日：2020-04-29

Applicant: Cytiva Bioprocess R&D AB

Inventor： Gustav Rodrigo ,  Mats Ander ,  Tomas Bjorkman ,  Ronnie Palmgren ,  Charlotte Brink

IPC: C07K16/12 ,  C07K1/22

Abstract: The invention discloses a method for separation of antibodies or antibody fragments, comprising the steps of: a) providing a feed comprising antibodies or antibody fragments having a VH3 region and being devoid of an Fc region capable of binding to Protein A; b) contacting the feed with a separation resin having covalently coupled ligands, wherein the ligands comprise a polypeptide as defined by SEQ ID NO 1 and wherein the antibodies or antibody fragments bind to the separation resin; c) optionally washing the separation resin with a washing liquid; d) eluting the antibodies or antibody fragments from the separation resin with an elution liquid and recovering the antibodies or antibody fragments.

公开(公告)号：US11124539B2

公开(公告)日：2021-09-21

申请号：US16348534

申请日：2017-11-14

Applicant: Cytiva BioProcess R&D AB

Inventor： Johan William Ohman ,  Mats Ander ,  Therese Graner ,  Anneli Jorsback ,  Jinyu Zou

IPC: C07K1/22 ,  B01J20/32 ,  B01D15/38 ,  B01J20/24 ,  B01J20/285 ,  B01J20/286

Abstract: The present invention relates to the field of chromatography and more specifically to producing protein affinity chromatography resins comprising affinity ligands based on a N-terminal fragment of a split intein, such as DnaE from Nostoc punctiforme, as well as methods for using the same. The N-terminal fragments are produced in inclusion bodies in bacterial cells.