Ge Healthcare Bio-Sciences Ab

Ge Healthcare Bioprocess R&d Ab

Henrik Ihre

Anders Larsson

Nils Norrman

Gunnar Glad

Andreas Axen

Hans Berg

Anders Hagvall

Dag Lindstrom

Eva Holmgren

David Buckley

Tobias Soderman

Jan Bergstrom

James Van Alstine

Philippe Busson

Andreas Axén

Maria Brännholm

Dag Lindström

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 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.

Method of preparing a separation matrix

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.

Matrix for separation of polyethers and method of separation

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.

Manufacture of a chromatography matrix

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.

Method of preparing a chromatography matrix

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.

Post-modification of a porous support

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.

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Uppsala, Sweden

Henrik Ihre