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.

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 separation matrix comprised of a porous or non-porous support to which ligands have been immobilised, wherein said ligands comprise at least one aliphatic sulfamide. The invention also relates to a chromatography column that contains the described separation matrix, as well as to a method of isolating immunoglobulins, such as IgG, Fab fragments, fusion proteins containing immunoglobulins etc, by adsorption to a separation matrix that comprises the aliphatic sulfamide ligands of the invention.

Abstract: The present invention relates to a method of separating antibodies from other compound(s) in a liquid sample, wherein a mobile phase comprising said sample is contacted with a multi-modal separation matrix to adsorb undesired compounds while the antibodies remain free in the liquid, wherein the multi-modal separation matrix comprises first groups, which are capable of interacting with negatively charged sites of the target compounds, and second groups, which are capable of at least one interaction other than charge-charge interaction with said target compounds. The invention also relates to a chromatography column packed with the above-described multi-modal separation matrix and a filter having such multi-modal groups adsorbed to its surface.

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 method of isolating target compounds from a liquid, which method including the steps of providing a mobile phase, which contains at least one target compound and wherein the conductivity corresponds to ?0.6 M; contacting the mobile phase with a separation matrix including one or more sulphonamide groups to adsorb one or more target compounds; contacting an eluent with the matrix to release target compound(s), wherein the conductivity of the eluent is reduced as compared to the mobile phase conductivity and the pH is substantially equivalent to the mobile phase pH; and, optionally, recovering at least one target compound or a purified liquid.

Abstract: The present invention relates to a method of separating antibodies from contaminants in a solution, which method comprises contacting the solution with a chromatography resin comprised of a support to which multi-modal ligands have been immobilised, wherein a multi-modal ligand comprises at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system. In one embodiment, the ring-forming atoms of the aromatic or hereoaromatic entity are selected among C, S or O, and the cation exchanging group is a weak cation exchanger. The present method may be used as a single step procedure or as a polishing step following a capture on a Protein A column.

Abstract: The present invention relates to a method of isolating fully thioated single stranded antisense oligonucleotides from a biological solution, which method comprises the steps of contacting the biological solution with an immobilised metal ion adsorption chromatography (IMAC) resin to adsorb the antisense oligonucleotides to the resin and subsequently contacting the resin with an eluent under conditions that provide desorption of the antisense oligonucleotides from the resin, wherein the fully thioated antisense oligonucleotides are separated from incorrectly thioated antisense oligonucleotides in the solution. The invention also relates to the use of an immobilised metal ion adsorption chromatography (IMAC) resin for isolation of fully thioated single stranded antisense oligonucleotides from a biological solution.

Abstract: The invention is a method of preparing multimodal ligands for hydrophobic interaction chromatography (HIC), which comprises providing a cyclic scaffold comprising a thiol, an amine and a carbonyl group; derivatisation of the nitrogen with a reagent to introduce a primary interaction; and aminolysis of the resulting derivative, whereby a secondary interaction is introduced next to the carbonyl; wherein at least one of the primary interaction and the secondary interaction comprises a hydrophobic group and wherein non of said interactions comprises charged groups, i.e. ion exchange ligands. The invention also encompasses a separation medium comprising such multimodal ligands immobilised to a base matrix.

Abstract: The present invention relates to a method of separating antibodies from other compound(s) in a liquid sample, wherein a mobile phase comprising said sample is contacted with a multi-modal separation matrix to adsorb undesired compounds while the antibodies remain free in the liquid, wherein the multi-modal separation matrix comprises first groups, which are capable of interacting with negatively charged sites of the target compounds, and second groups, which are capable of at least one interaction other than charge-charge interaction with said target compounds.

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 comprised of a porous support to which ligands have been immobilised, 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 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.