Abstract: The present invention relates to a method of producing novel chromatography media and use thereof for purification of biomolecules, such as proteins. The chromatography media comprises shell beads having an inner porous core and an outer shell. The method comprises providing buffering ligands in the core of the beads, and providing binding ligands aimed for biomolecule binding in the outer shell of the beads. This method makes it possible to optimize binding properties and buffering properties independently of each other which is especially to advantageous for production of chromatofocusing media.

Abstract: The present invention relates to a buffering composition, more closely a composition comprising porous matrix-enclosed buffering agent(s) giving a stabilisation of pH when applied in for example aqueous environment. The composition comprises buffering agent(s) enclosed in a first porous matrix with an impenetrability corresponding to a fractionation range for globular proteins and peptides of <10000 g/mol, preferably <5000 g/mol, more preferably <700 g/mol.

Abstract: The present invention relates to a method for running ion exchange chromatography on a media comprising shell beads having an inner porous core and an outer shell, wherein the inner core is provided with ligands whose charge changes with pH and the shell is provided with charged ion exchange ligands, the method comprising the following steps: a) adsorbing sample molecules on the shell ligands at a first pH; b) causing a discharge of the inner core ligands at a second pH by addition of a buffer substance that is able to increase its charge having the same sign/type as that of the core ligands, which at the same time causes release of ions from the inner core ligands and thereby an increase in ionic strength that displaces the sample molecules from the shell ligands i.e. causes an elution.

Abstract: The present invention relates to a method and means to produce chromatography media having improved pressure-flow properties. More closely, the invention relates to bimodal particle size distribution and the use of layer functionalisation as means to change pressure-flow properties of chromatography media. The invention relates to a method for production of chromatography media having improved pressure-flow properties, comprising mixing large beads/particles, comprising an inner core and an outer functionalized shell/lid, with smaller beads/particles, wherein the ratio of the particle size of large and small beads: [D50V for large particles/D50V for small particles]>1.2, and wherein the volume ratio of large and small beads in the column: [Total volume of large beds/Total volume beads] is in the range 0.05-0.9.

Abstract: 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° 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° C., preferably between 17.5 and 24.6° 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.

Abstract: The present invention relates to a method of producing novel chromatography media and use thereof for purification of biomolecules, such as proteins. The chromatography media comprises shell beads having an inner porous core and an outer shell. The method comprises providing buffering ligands in the core of the beads, and providing binding ligands aimed for biomolecule binding in the outer shell of the beads. This method makes it possible to optimize binding properties and buffering properties independently of each other which is especially to advantageous for production of chromatofocusing media.

Abstract: 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.

Abstract: The present invention relates to a method of preparing a library of resins which are useful in chromatography, which method comprises creating a diversity of multi-modal ion exchange resins; and providing the diversity in a parallel system in which each resin is presented separated from the other resin(s).

Abstract: The present invention relates to a separation matrix comprised of ligands coupled to the surfaces of a porous support, such as one or more porous particles, wherein the ligands provide at least one chemical gradient within the support. In the most advantageous embodiment, the chemical gradient is a ligand density gradient. The invention also relates to a method of providing a separation matrix comprising ligands coupled to the surfaces of a porous support, in which method at least one ligand density gradient is provided by solvent-controlled diffusion of at least one reagent into the porous support.

Abstract: The present invention relates to a chromatography ligand defined by the following formula R1—R2—N(R3)—R4—R5 wherein R1 is a substituted or non-substituted phenyl group; R2 is a hydrocarbon chain comprising 0-4 carbon atoms; R3 is a hydrocarbon chain comprising 1-3 carbon atoms; R4 is a hydrocarbon chain comprising 1-5 carbon atoms; and R5 is 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.

Abstract: 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.

Abstract: The present invention relates to a chromatography ligand defined by the following formula R1—R2—N(R3)—R4—R5 wherein R1 is a substituted or non-substituted phenyl group; R2 is a hydrocarbon chain comprising 0-4 carbon atoms; R3 is a hydrocarbon chain comprising 1-3 carbon atoms; R4 is a hydrocarbon chain comprising 1-5 carbon atoms; and R5 is 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.

Abstract: The present invention relates to a method for removing at least one negatively charged substance from an aqueous liquid by contacting the liquid with a separation matrix comprising a plurality of polyallylamine ligands, comprising binding said negatively charged substance to said ligands under conditions where the ionic strength of the aqueous liquid applied to the chromatography resin ?0.25 M NaCl.

Abstract: 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.

Abstract: A method for the removal of a substance, which has a negative charge and which is present in an aqueous liquid (I). The method comprises the steps of: (i) contacting the liquid with an anion-exchanger (1) that comprises mixed mode anion-exchanging ligands in which there is a positively charged nitrogen allowing binding of the substance to the anion-exchanger; and (ii) desorbing said substance from said anion-exchanger. The characteristic feature is that (A) the mixed mode ligands have a thioether linkage within a distance of 1-7 atoms from their positively charged atom, and (B) the anion-exchanger (1) (i) is capable of binding the substance of interest in an aqueous reference liquid (II) at an ionic strength corresponding to 0.25 M NaCl, and (ii) permits in the pH interval 2-12 a maximal breakthrough capacity for the substance which is ?200% of the breakthrough capacity of the substance for Q-Sepharose Fast Flow (anion-exchanger 2).

Abstract: The present invention relates to a chromatography ligand defined by the following formula (I) R1—R2—N(R3)—R4—R5 wherein R1 is a substituted or non-substituted phenyl group; R2 is a hydrocarbon chain comprising 0-4 carbon atoms; R3 is a hydrocarbon chain comprising 1-3 carbon atoms; R4 is a hydrocarbon chain comprising 1-5 carbon atoms; and R5 is 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.

Abstract: 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.

Abstract: 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° 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° C., preferably between 17.5 and 24.6° 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.

Abstract: The present invention relates to a process for the purification of antibodies from one or more impurities in a liquid, which process comprises contacting said liquid with a first chromatography resin comprised of a support to which multi-modal ligands have been immobilised to adsorb the antibodies to the resin, wherein each multi-modal ligand comprises at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system; adding an eluent to release the antibodies from the resin; and contacting the eluate so obtained with a second chromatography resin. In one embodiment, the ring-forming atoms of the aromatic or heteroaromatic entity are selected from the group consisting of C, S and O, and the cation exchanging group is a weak cation exchanger.

Abstract: The present invention relates to a separation matrix comprised of a porous support to which ligands have been immobilised, 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.