Ge Healthcare Bio-sciences Ab

Ge Healthcare Bioprocess R&d Ab

Amersham Biosciences Ab

Cytiva Bioprocess R&d Ab

Bo-Lennart Johansson

Jean-Luc Maloisel

Gunnar Glad

Hans J Johansson

Nils Norrman

Jan Bergstrom

Annika Forss

Makonnen Belew

Carina Engstrand

Nicolas Thevenin

Tobias Soderman

Kjell Eriksson

Enrique Napoleon Carredano

Andreas Axen

Jamil Shanagar

Anna Grönberg

Eggert Brekkan

Anders Ljunglof

Jafar Saidzadeh

Ake Stenholm

Anders Ljunglöf

Mikael Andersson

Jan Gustavsson

Asa Ek

Maria Holm

Jean-Luc Ge Healthcare Maloisel

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 is within the field of biomolecule purification. More closely the invention relates to chromatographic purification of insulin using a specific kind of shell beads having an inner core and an outer functionalized layer. The method enables purification at high flow rates and high purity, over 90%.

Method for purification of cleaved pro-insulin

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.

Method for ion-exchange chromatography and media used thereof

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 functionalization 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: [Dfor large particles/Dfor small particles]&#x3e;1.2, and wherein the volume ratio of large and small beads in the column: [Total volume of large beads/Total volume beads] is in the range 0.05-0.9.

Method for production of chromatography media

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 separation matrix comprised of a porous or non-porous support to which ligands have been immobilized, 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.

Purification of immunoglobulins

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.

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.

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.

Growing community of inventors

Uppsala, Sweden

Bo-Lennart Johansson