公开(公告)号：US11667671B2

公开(公告)日：2023-06-06

申请号：US16911511

申请日：2020-06-25

Applicant: Cytiva BioProcess R&D AB

Inventor： Annika Forss ,  Mats Ander ,  Tomas Bjorkman ,  Hans Blom ,  Jesper Hansson ,  Gustav Rodrigo

IPC: C07K1/16 ,  C07K1/22 ,  C07K16/00 ,  B01J20/286 ,  B01J20/285 ,  B01D15/18 ,  B01D15/34 ,  C07K14/31

CPC classification number: C07K1/165 ,  B01D15/1807 ,  B01D15/1864 ,  B01D15/34 ,  B01J20/285 ,  B01J20/286 ,  C07K1/22 ,  C07K14/31 ,  C07K16/00 ,  C07K2317/21 ,  C07K2317/31 ,  C07K2319/30

Abstract: The invention relates to a method of isolating an immunoglobulin, comprising the steps of:



a) providing a separation matrix comprising at least 15 mg/ml multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support, wherein 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;
b) contacting a liquid sample comprising an immunoglobulin with the separation matrix;
c) washing the 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 comprising at least 0.5 M NaOH.

公开(公告)号：US20250066757A1

公开(公告)日：2025-02-27

申请号：US18719120

申请日：2023-02-20

Applicant: Cytiva BioProcess R&D AB

Inventor： Simon Aberg ,  Linda Hagman ,  Mikael Berg ,  Hans Blom ,  Albin Larsson ,  Joakim Bagge ,  Elin Monie ,  Markus Pitkanen

IPC: C12N15/10

Abstract: The present disclosure is directed to a method for separating supercoiled plasmid DNA (pDNA) from a liquid sample. the method comprising the steps of: (a) adding a liquid sample comprising pDNA to a first chromatography material comprising (i) an anion exchange chromatography ligand for binding to pDNA and (ii) a support material allowing convective flow through the first chromatography material, wherein the liquid sample originates from a cell culture harvest and has been subjected to a step of removing RNA before step (a): (b) eluting a liquid sample. comprising a purified mixture of supercoiled pDNA and open circular pDNA, from the first chromatography material: (c) adding the liquid sample from step (b) to a second chromatography material comprising a ligand that binds to pDNA and enables selective separation of supercoiled pDNA from open circular pDNA: (d) eluting the purified supercoiled pDNA from the second chromatography material: wherein the supercoiled pDNA eluted in step (d) has a purity degree of at least 95% without use of any further chromatography material than said first and second chromatography materials. Steps (a)-(d) and any intermediate steps can be completed within 5 hours. Further disclosed are uses of supercoiled pDNA obtained by said separation method.

公开(公告)号：US20220111310A1

公开(公告)日：2022-04-14

申请号：US17544700

申请日：2021-12-07

Applicant: Cytiva Bioprocess R&D AB

Inventor： Karol Maciej Lacki ,  Hans Blom ,  Helena Skoglar ,  Linus Laurin

IPC: B01D15/38 ,  B01J20/24 ,  B01J20/286 ,  B01J20/32 ,  C07K1/22 ,  B01J20/28 ,  C07K16/06

Abstract: The invention discloses a separation matrix comprised of porous spherical particles to which antibody-binding protein ligands have been covalently immobilized, wherein the density of said ligands is in the range of 10.5-15 mg/ml and the volume-weighted median diameter of said particles is in the range of 30-55 μm.