Ge Healthcare Bio-sciences Ab

Cytiva Bioprocess R&d Ab

Ge Healthcare Bioprocess R&d Ab

Ge Healthcare Bioprocess R&d

Per-Mikael Aberg

Gustav Jose Rodrigo

Martin Hall

Jinyu Zou

Andreas Muranyi

Sture Larsson

Tomas Bjorkman

Mats Ander

Hans J Johansson

Hans Berg

Bjorn Noren

Goran Bauren

Gunnar Malmquist

Göran Baurén

Tomas Björkman

Jelena Vasic

Gustav Rodrigo

The present invention relates to a chromatography ligand, which comprises Domain C fromprotein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIP) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates to a process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.

Chromatography ligand comprising domain c fromprotein a for antibody isolation

The present invention relates to a chromatography ligand, which comprises Domain C fromprotein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIF) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.

Chromatography ligand comprising domain C fromprotein A for antibody isolation

The present invention relates to a chromatography ligand, which comprises Domain C fromprotein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIP) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.

[object Object]

The invention discloses an immunoglobulin-binding protein comprising one or more mutated immunoglobulin-binding domains (monomers) of staphylococcal Protein A (E, D, A, B, C) or protein Z or a functional variant thereof, wherein in at least one of the one or more mutated monomers, the asparagine or histidine at the position corresponding to H18 of the B domain of Protein A or of Protein Z has been deleted or substituted with a first amino acid residue which is not proline or asparagine and wherein, if the amino acid residue at position 57 is proline and the amino acid residue at position 28 is asparagine, then the amino acid residue at the position corresponding to H18 of the B domain of protein A or of protein Z is not serine, threonine or lysine.

Affinity chromatography matrix

The present invention relates to a method of separating one or more immunoglobulin containing proteins from a liquid. The method includes first contacting the liquid with a separation matrix comprising ligands immobilized to a support; allowing the immunoglobulin containing proteins to adsorb to the matrix by interaction with the ligands; followed by an optional step of washing the matrix containing the immunoglobulin containing proteins adsorbed thereon; and recovering said immunoglobulin containing proteins by contacting the matrix with an eluent which releases the proteins. The method improves upon previous separation methods in that each of the ligands comprises one or more of a protein A domain (E, D, A, B, C), or protein Z, or a functional variant thereof, with at least one of the monomers having a substitution of the Asparagine or Histidine at the position corresponding to H18 of B domain of Protein A or Protein Z, and wherein the ligand provides an increase in elution pH compared to non-substituted ligand.

The present invention relates to a separation matrix comprised of porous particles to which antibody-binding protein ligands have been immobilized, wherein the ligand density is in the range of 5.0-10 mg/ml; the gel phase distribution coefficient of the particles expressed as Kfor a dextran of size 110 kDa is above 0.65 and the median particle diameter is between 65-84 &#x3bc;m. The carbohydrate material is preferably highly cross-linked agarose.

Purification of immunoglobulins

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Uppsala, Sweden

Per-Mikael Aberg