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    Method of Storing a Separation Matrix
    4.
    发明申请
    Method of Storing a Separation Matrix 审中-公开

    公开(公告)号：US20200239517A1

    公开(公告)日：2020-07-30

    申请号：US16095721

    申请日：2017-05-10

    Applicant: GE HEALTHCARE BIOPROCESS R&D AB

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

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

    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.

    Separation method
    5.
    发明授权
    Separation method 有权

    公开(公告)号：US10703774B2

    公开(公告)日：2020-07-07

    申请号：US15348699

    申请日：2016-11-10

    Applicant: GE Healthcare BioProcess R&D AB

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

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

    Method of Cleaning and/or Sanitizing a Separation Matrix
    7.
    发明申请
    Method of Cleaning and/or Sanitizing a Separation Matrix 审中-公开

    公开(公告)号：US20200299325A1

    公开(公告)日：2020-09-24

    申请号：US16095753

    申请日：2017-05-10

    Applicant: GE HEALTHCARE BIOPROCESS R&D AB

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

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

    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.

    Separation method
    8.
    发明授权
    Separation method 有权

    公开(公告)号：US10730908B2

    公开(公告)日：2020-08-04

    申请号：US15798784

    申请日：2017-10-31

    Applicant: GE HEALTHCARE BIOPROCESS R&D AB

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

    IPC: C07K1/16 ,  B01D15/08 ,  C07K1/22 ,  C07K16/00 ,  B01J20/286 ,  B01J20/285 ,  B01D15/18 ,  B01D15/34 ,  C07K14/31

    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.

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