Ge Healthcare Bio-sciences Ab

Ge Healthcare Bioprocess R&d Ab

Cytiva Bioprocess R&d Ab

Gunnar Glad

Jean-Luc Maloisel

Bo-Lennart Johansson

Nils Norrman

Hans J Johansson

Annika Forss

Carina Engstrand

Jan Bergstrom

Andreas Axen

Tobias Soderman

Anders Larsson

Nicolas Thevenin

Henrik Ihre

Mathias Alterman

Dag Lindstrom

Andreas Axén

Therese Graner

Susanna Lindman

Ulrika Meyer

Anders Ljunglof

Ake Stenholm

The present invention is within the field of chromatography. More precisely, it relates to a novel chromatography medium, namely a hydrophobic medium provided with different lids excluding molecules over a certain size due to the porosity of the hydrophobic medium and/or the porosity of the lid. The invention also relates to use of the separation medium for purification of large molecules, which do not enter the separation medium, as well as small molecules, which enter the separation medium and are eluted from there.

Chromatography medium

The present invention relates to a chromatography ligand defined by the following formula R&#x2014;R&#x2014;N(R)&#x2014;R&#x2014;Rwherein Ris a substituted or non-substituted phenyl group; Ris a hydrocarbon chain comprising 0-4 carbon atoms; Ris a hydrocarbon chain comprising 1-3 carbon atoms; Ris a hydrocarbon chain comprising 1-5 carbon atoms; and Ris OH or H. The invention also comprises a separation matrix, comprising the described ligands coupled to a porous support, such as particles or a membrane. The ligand and matrix according to the invention is useful for purification of biomolecules or organic compounds, such as proteins, polypeptides, DNA etc. An advantageous use according to the invention is the purification of antibodies.

Chromatography ligand

The present invention relates to a separation matrix, which comprises a support; extenders coupled to an outer part of said support; and ligands coupled to said extenders, wherein the part of the support to which the extenders are coupled constitutes less than 50% of the volume of the separation matrix. The invention also embraces a method of preparing such a separation matrix, as well as a process wherein the separation media is used.

Separation matrix

The present invention relates to a novel non-cross linked chromatography media provided with glutathione ligands which may or may not be provided with magnetic particles. The chromatography media is used for production of an affinity media provided with gluthatione ligands for adsorption of GST-tagged proteins.

Chromatography media and use thereof

The present invention relates to a separation medium, comprising an inner core of a porous material provided with charged ligands, and an outer lid comprising a porous material provided with charged ligands, wherein the charge of the ligands in the inner core is opposite that of the charge of the ligands in the lid. The present invention also relates to a method for biomolecule separation comprising applying a sample to the above separation medium, wherein large molecules are prevented from entering the medium by charge repulsion from the medium and small molecules are captured in the inner core.

Separation medium for chromatography of various biomolecules

The present invention relates to a method of manufacturing a chromatography matrix comprising providing a polysaccharide carrier comprising available hydroxyl groups; and reacting said hydroxyl groups with vinyl sulphonate to provide a sulphonate-functionalized (S-functionalized) cation exchanger. The hydroxyl groups of the carrier may be hydroxyls of the agarose polymer; or alternatively they may be provided on extenders such as polyhydroxyfunctional polymers. In one embodiment, the carrier is made of agarose with improved flow pressure properties.

Manufacture of chromatography matrices

The present invention relates to a separation matrix comprised of a porous support to which ligands have been immobilized, wherein said ligands comprise at least one aliphatic sulphonamide. The nitrogen of the sulphonamide may be a secondary or tertiary amine. The invention also relates to a chromatography column that contains the described separation matrix, as well as to a method of isolating immunoglobulin-like compounds by adsorption to a separation matrix that comprises aliphatic sulphonamide ligands.

Purification of immunoglobulins

Disclosed is a method for activating a solid support material with epoxy groups and for immobilizing ligands thereon, utilizing phase transfer catalytic conditions. The method permits the introduction of epoxy groups and specific nucleophilic ligands on the support material with a high level of substitution. Furthermore, the invention provides a general method for immobilizing a ligand for use in a wide variety of chromatographic separation procedures such as ion exchange chromatography, hydrophobic interaction chromatography (HIC), reverse phase chromatography (RPC), or affinity chromatography.

Activated solid support and method

The present invention relates to a method for sample application and separation. More closely, the invention relates to convenient direct loading of a biomolecule sample via magnetic beads to, for example, a gel before electrophoresis. In this way, the invention combines elution and application steps with minimal losses of sample. Thus, the invention relates to a method for sample application of biomolecules on a separation media, comprising the following steps: a) obtaining said biomolecules from a sample by magnetic beads; b) applying the magnetic beads with the biomolecules to a separation medium; c) releasing the biomolecules into the separation media, and d) separation of the biomolecules from each other in the separation medium.

Method for sample application

The present invention relates to a method for production of separation media using a so called Spinning Disc technology wherein the porosities of the beads are optimized in such a way that a desired biomolecule may be separated from a complex sample. The method comprises the following steps: a) feeding a 4-8% polysaccharide solution, which has a viscosity within 350-450 mPas, at 65-75&#xb0; C. to one or more spinning discs at 3001-3010 rpm to form polysaccharide beads; b) capturing said formed polysaccharide beads in a capturing bath; wherein the porosity of the polysaccharide beads is controlled by varying the temperature of the capturing between 15 and 27&#xb0; C., preferably between 17.5 and 24.6&#xb0; C. The method yields porosities that prevent molecules larger than 150 000 g/mol to diffuse into the beads. The invention also relates to separation media produced by the method and use thereof for purification of biomolecules, in particular monoclonal antibodies.

Method for production of separation media

The present invention relates to a separation matrix comprised of a porous support to which ligands have been immobilized, wherein said ligands comprise at least one sulphonamide and the R group of the sulphonyl comprises an aromatic group. The nitrogen of the sulphonamide may be a secondary or tertiary amine. The invention also relates to a chromatography column that contains the described separation matrix, as well as to a method of isolating immunoglobulin-like compounds by adsorption to the separation matrix.

Growing community of inventors

Uppsala, Sweden

Gunnar Glad