Abstract: 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.

Abstract: The present invention relates to a method of preparing an insoluble separation matrix, which method comprises salt-treatment of a polysaccharide gel followed by cross-linking of the polysaccharide polymers. In one embodiment, the method comprises providing an aqueous solution of a gelatable native polysaccharide; lowering the temperature of the polysaccharide solution to a value below its gelling point; salt-treatment by adding at least one salt to the resulting polysaccharide; and cross-linking the salt-treated polysaccharide. The polysaccharide may be prepared e.g. into particles, membranes or monoliths.

Abstract: The present invention relates to a separation matrix comprised of a support to the surfaces of which polymer chains have been coupled, wherein each polymer chain presents recurring proton-donating groups and at least the surface of the support is substantially hydrophilic. In the most advantageous embodiment, the support is porous cross-linked agarose, the polymers are poly(acrylic acid) and the proton-donating groups are carboxyl groups. The matrix is useful e.g. to remove PEG from pegylated and/or native compounds in a liquid. Accordingly, the invention also encompasses a method, such as a chromatographic method, wherein the separation matrix according to the invention is used, for example as a pre-treatment of a reaction mixture that comprises unreacted PEG, pegylated proteins and native proteins.

Abstract: The present invention relates to a separation matrix comprised of a support to the surfaces of which polymer chains have been coupled, wherein each polymer chain presents recurring proton-donating groups and at least the surface of the support is substantially hydrophilic. In the most advantageous embodiment, the support is porous cross-linked agarose, the polymers are poly(acrylic acid) and the proton-donating groups are carboxyl groups. The matrix is useful e.g. to remove PEG from pegylated and/or native compounds in a liquid. Accordingly, the invention also encompasses a method, such as a chromatographic method, wherein the separation matrix according to the invention is used, for example as a pre-treatment of a reaction mixture that comprises unreacted PEG, pegylated proteins and native proteins.

Abstract: The present invention relates to method of manufacturing a sulphonate-functionalized (S-functionalized) cation exchanger, which method comprises reacting olefinic groups with bisulphite in the presence of at least one amine oxide, such as N-methyl morpholine oxide (NMO). The olefinic groups of the carrier may be allyl groups; which may be provided on extenders such as polyhydroxyfunctional polymers. In one embodiment, the carrier is made of agarose with improved flow pressure properties.

Abstract: The present invention relates to method of manufacturing a sulphonate-functionalized (S-functionalized) cation exchanger, which method comprises reacting olefinic groups with bisulphite in the presence of at least one amine oxide, such as N-methyl morpholine oxide (NMO). The olefinic groups of the carrier may be allyl groups; which may be provided on extenders such as polyhydroxyfunctional polymers. In one embodiment, the carrier is made of agarose with improved flow pressure properties.

Abstract: 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.

Abstract: The present invention relates to a method of preparing a porous cross-linked polysaccharide chromatography matrix, which comprises to provide an aqueous solution of a gellable polysaccharide, wherein part of the hydroxyl groups are substituted with groups which are not susceptible to nucleophilic attack; to provide essentially spherical droplets of the substituted polysaccharide solution; to form a gel of the substituted polysaccharide solution; and to cross-link the gel by adding a cross-linking agent. The invention also encompasses a chromatography column packed with a matrix so produced as well as use thereof in e.g. protein purification.

Abstract: The present invention relates to a method of preparing an insoluble separation matrix, which method comprises salt-treatment of a polysaccharide gel followed by cross-linking of the polysaccharide polymers. In one embodiment, the method comprises providing an aqueous solution of a gelatable native polysaccharide; lowering the temperature of the polysaccharide solution to a value below its gelling point; salt-treatment by adding at least one salt to the resulting polysaccharide; and cross-linking the salt-treated polysaccharide. The polysaccharide may be prepared e.g. into particles, membranes or monoliths.

Abstract: The present invention relates to a separation matrix comprised of a support to the surfaces of which polymer chains have been coupled, wherein each polymer chain presents recurring proton-donating groups and at least the surface of the support is substantially hydrophilic. In the most advantageous embodiment, the support is porous cross-linked agarose, the polymers are poly(acrylic acid) and the proton-donating groups are carboxyl groups. The matrix is useful e.g. to remove PEG from pegylated and/or native compounds in a liquid. Accordingly, the invention also encompasses a method, such as a chromatographic method, wherein the separation matrix according to the invention is used, for example as a pre-treatment of a reaction mixture that comprises unreacted PEG, pegylated proteins and native proteins.

Abstract: The present invention relates to a method for post-modification of a macroporous polymeric support comprising one or more unreacted double bonds, which method includes the steps of contacting the support with a liquid comprising a grafting compound; and initiating a free radical reaction in the mixture obtained, wherein the grafting compound is comprised of a double bond as a reactive group coupled to a linear or cyclic chain of carbon. The grafting compound can be illustrated with vinyl ethers and/or styrene and styrene derivatives.

Abstract: The present invention relates to a method for post-modification of a porous polymeric polysaccharide support comprising one or more unreacted double bonds, which method includes to contact the polymeric support with a liquid phase comprising at least one grafting compound and to initiate a free radical reaction in the mixture obtained, wherein the grafting compound is comprised of a double bond as a reactive group coupled to a linear or cyclic chain of carbon, which optionally comprises one or more heteroatoms. An advantageous grafting compound is a vinyl ether or a hydroxyvinyl ether. The invention also encompasses a polymeric support that has been post-modified by the method according to the invention as well as the use of such a support, e.g. in chromatography.

Abstract: The present invention relates to a method of preparing a porous cross-linked polysaccharide chromatography matrix, which comprises to provide an aqueous solution of a gellable polysaccharide, wherein part of the hydroxyl groups are substituted with groups which are not susceptible to nucleophilic attack; to provide essentially spherical droplets of the substituted polysaccharide solution; to form a gel of the substituted polysaccharide solution; and to cross-link the gel by adding a cross-linking agent. The invention also encompasses a chromatography column packed with a matrix so produced as well as use thereof in e.g. protein purification.

Abstract: The present invention relates to a method for post-modification of a macroporous polymeric support comprising one or more unreacted double bonds, which method includes the steps of contacting the support with a liquid comprising a grafting compound; and initiating a free radical reaction in the mixture obtained, wherein the grafting compound is comprised of a double bond as a reactive group coupled to a linear or cyclic chain of carbon. The grafting compound can be illustrated with vinyl ethers and/or styrene and styrene derivatives.

Abstract: The present invention relates to a method for post-modification of a porous polymeric polysaccharide support comprising one or more unreacted double bonds, which method includes to contact the polymeric support with a liquid phase comprising at least one grafting compound and to initiate a free radical reaction in the mixture obtained, wherein the grafting compound is comprised of a double bond as a reactive group coupled to a linear or cyclic chain of carbon, which optionally comprises one or more heteroatoms. An advantageous grafting compound is a vinyl ether or a hydroxyvinyl ether. The invention also encompasses a polymeric support that has been post-modified by the method according to the invention as well as the use of such a support, e.g. in chromatography.