Ge Healthcare Bio-sciences Ab

Ge Healthcare Bioprocess R&d Ab

Nicolas Thevenin

Jean-Luc Maloisel

Bo-Lennart Johansson

Gunnar Glad

Hans J Johansson

Anders Larsson

Andreas Axen

Mattias Algotsson

Philippe Busson

Anders Hagvall

Anna Grönberg

Anders Ljunglöf

Ulrika Carlsson

Disclosed is a method for activating a solid support material with epoxy groups and for immobilizing ligands thereon, utilizing phase transfer catalytic conditions. The method permits the introduction of epoxy groups and specific nucleophilic ligands on the support material with a high level of substitution. Furthermore, the invention provides a general method for immobilizing a ligand for use in a wide variety of chromatographic separation procedures such as ion exchange chromatography, hydrophobic interaction chromatography (HIC), reverse phase chromatography (RPC), or affinity chromatography.

Activated solid support and method

The invention relates to a process for making porous cross-linked cellulose membranes and processes for coupling a chromatography ligand to cross-linked cellulose membranes. The invention provides methods for separating a first component from a second component in a solution based upon a difference in the size of the first and second components, and methods for separating target molecules from other components in a solution comprising use of membranes obtainable by the process of the invention. The method has particular utility in separating proteins from cell lysates and cultures.

Process for cross-linking cellulose ester membranes

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.

Process for the purification of antibodies

The present invention is a method of preparing affinity ligands, which method comprises to provide a cyclic scaffold comprising a thiol group, a carbonyl group and an amine group; to derivatize the amine group of said scaffold with an electrophile that carries either a functionality capable of affinity interaction or a functionality capable of a second interaction with a target molecule; and to open up the derivatized scaffold and add an amine that carries either a functionality capable of a second interaction or a functionality capable of affinity interaction with a target molecule. The method provides provide affinity ligands, which are capable of affinity interaction as well as a second kind of interaction.

Method of preparing affinity ligands

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.

Antibody purification

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.

Method of preparing ligands for hydrophobic interaction chromatography

The invention relates to a method of preparing multi-modal anion-exchange ligands, which comprises providing a cyclic three-functional scaffold comprising an amine, a carbonyl and a thiol; optionally derivatisation of the amine of the scaffold to provide an anion-exchanging group and/or to protect the amine; and aminolysis to open up the derivative by adding a reagent comprising an amine coupled to a residue R in order to add said amine to the carbonyl carbon of the opened scaffold. The scaffold is advantageously homocysteine thiolactone. In one embodiment, the method comprises an additional step of immobilising the opened scaffold to a base matrix to provide a separation medium, such as a chromatography medium.

Method of preparing multi-modal anion-exchange ligands

The present invention relates to a method for synthesis of an acrylamide derivative, starting with dissolving a salt of a nucleophilic amine in water to form an aqueous solution and desalting said solution with a base, comprising the following steps:

Method for synthesis of acrylamide derivatives

The present invention relates to a method of generating a separation medium comprising mixed mode cation-exchanger ligands coupled to a base matrix, which method comprises to provide a scaffold comprising a functional group and exhibiting a cyclic core structure; derivatise the scaffold with a reagent comprising a reactive group coupled to a residue R by reacting the functional group of the scaffold with said reactive group; open the cyclic structure of the resulting derivative; and react the product with a base matrix comprising a reactive group. The scaffold presents at least two functionalities; one sulphur-comprising group for coupling to the base matrix and one group that can be transformed into an ionic group.

Growing community of inventors

Uppsala, Sweden

Nicolas Thevenin