Abstract: The invention relates to a process for making porous cross-linked charged cellulosic polymeric membranes capable of binding to a target molecule. The invention provides 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 and nucleic acids from cell lysates and cultures.

Abstract: The present invention relates to a method and a device for small scale reactions, such as sample preparation of a desired substance in a sample. In the method using the device samples mixed with functionalized magnetic particles are magnetically transferred between different working stations on the device. The method uses a hydrophobic surface, such as a Petri dish, provided with hydrophilic spots of, for example, agarose beads located on said hydrophobic surface and provided with buffers, reactants or ligands.

Abstract: The present invention relates to an IgG-binding compound, which more specifically has affinity for human IgGs of ?-type and functional derivatives thereof. More specifically, the compound according to the invention comprises an N,N-alkylated urea moiety located between an aromatic part and another part, which is a linear or cyclic substituted or unsubstituted aliphatic group. The compound binds to a pocket-shaped binding site present on all human IgG ?-Fabs, which site is located between the two domains (CH1 and CL) of its constant part. Accordingly, the compound according to the invention is a ligand for human IgGs of ?-type, and consequently, the invention also relates to a separation matrix for affinity chromatography, which matrix comprises said compound, as well as to other uses of the compound.

Abstract: The present invention relates to a chromatographic adsorbent for selectively adsorbing IgG, comprising the following formula and its corresponding enol-form, wherein X represents O, S, or NH; R1 represents H, C1-6 alkyl, C1-6 alkoxy, C1-6 alkoxy-C1-6 alkyl, Ar, —C(O)NHR3, —C(O)—R3 or halo; R2 represents H, C1-3 alkyl or halo; R3 represents H, C1-6 alkyl, C1-6 alkoxy, C1-6 alkoxy-C1-6 alkyl or Ar; n represents 0, 1, 2 or 3; Y represents a carrier. The present invention also relates to a method of producing said adsorbent as well as use thereof for separating substances by affinity chromatography.

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 chromatographic adsorbent for selectively adsorbing IgG, comprising the following formula and its corresponding enol-form, wherein X represents O, S, or NH; R1 represents H, C1-6 alkyl, C1-6 alkoxy, C1-6 alkoxy-C1-6 alkyl, Ar, —C(O)NHR3, —C(O)—R3 or halo; R2 represents H, C1-3 alkyl or halo; R3 represents H, C1-6 alkyl, C1-6 alkoxy, C1-6 alkoxy-C1-6 alkyl or Ar; n represents 0, 1, 2 or 3; Y represents a carrier. The present invention also relates to a method of producing said adsorbent as well as use thereof for separating substances by affinity chromatography.

Abstract: The present invention relates to a method for generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of (a) providing at least one scaffold defined by the general formula (I): H2N—(X1)n—S—S—(X2)m—CH2—NH2 wherein X1 and X2 irrespective of each other are carbon atoms or heteroatoms, and n and m irrespective of each other are integers of 1 to 5; (b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities of at least one arm are protected, on each scaffold by derivatisation of the nucleophilic NH2 groups of the scaffold; (c) reducing the disulfide bond of the derivatised scaffold; and, if required (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment, the reduction of the disulfide bond and the deprotection step is performed essentially simultaneously.

Abstract: The present invention relates to a human IgG binding pocket comprised of a first interacting surface, which originates from an IgG ? light chain, and a second interacting surface, which originates from an IgG heavy chain, which amino acids are strictly conserved between human IgGs of ?-type. The invention also embraces an isolated and purified polypeptide, which comprises said binding pocket. Further, the invention relates to various methods of using the novel binding pocket, such as in screening for identification of chemical entities capable of selective binding thereof, and in other experimental and/or virtual methods for design and/or identification of chemical entities capable of selective binding thereof.

Abstract: The present invention relates to magnetic beads in the form of composite beads with an inner core of metal particles, which are coated with an inert synthetic polymer and thereafter a hydrophilic porous polymer, preferably dextran. This provides porous biocompatible beads without metal leakage. This construction also allows for simple and convenient handling of cell expansion media by magnetism.

Abstract: The present invention relates to a separation medium with various functionalities suitable for, for example, isolation of proteins, cells, and viruses and also for diagnostic applications and cell cultivation. The separation medium comprises magnetic metal particles, preferably coated with an inert synthetic polymer, and pre-functionalised beads. These particles and beads are provided encapsulated in a hydrophilic porous polymer, preferably agarose. The beads may be used for cell cultivation or for chromatography. When the beads are used for chromatography the agarose layer may be provided with ligands having affinity for selected biomolecules.

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 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.

Abstract: The present invention relates to magnetic beads suitable for, for example, isolation of proteins, cells, and viruses and also for diagnostic applications and cell cultivation. The magnetic beads are composite beads with an inner core of metal particles, which are coated with an inert synthetic polymer and these are then enclosed in a hydrophilic porous polymer, preferably agarose. This provides porous biocompatible beads without metal leakage. The beads may be used for cell cultivation or for chromatography. When the beads are used for chromatography the agarose layer is preferably provided with ligands having affinity for selected biomolecules.

Abstract: The present invention relates to the preparation of a metal chelating separation medium, which comprises providing a compound defined by formula: R—O—CO—(CH2)n—CHNH2—CO—OH wherein R is a selectively cleavable group; and n is an integer of 1–4; reacting it with ethyl alcohol; derivatisation; cleavage of R; immobilisation of the resulting compound to a base matrix; and deprotection of the carboxylic acid. In one embodiment, the compound provided in the first step is ethyl ester of aspartic acid or of glutamic acid, and the derivatisation agent is bromo-acetic acid ethyl acetate.

Abstract: The present invention relates to the preparation of a metal chelating separation medium, which comprises providing a compound defined by formula: R—O—CO—(CH2)n—CHNH2—CO—OH wherein R is a selectively cleavable group; and n is an integer of 1-4; reacting it with ethyl alcohol; derivatisation; cleavage of R; immobilisation of the resulting compound to a base matrix; and deprotection of the carboxylic acid. In one embodiment, the compound provided in the first step is ethyl ester of aspartic acid or of glutamic acid, and the derivatisation agent is bromo-acetic acid ethyl acetate.

Abstract: The present invention relates to a method of generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of a) providing at least one cyclic scaffold comprising a carbonyl, an adjacent sulphur and a nucleophile; b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities are protected, on each scaffold by derivatisation of the nucleophile of said scaffolds, while retaining the cyclic structure of the scaffold; (c) ring-opening at the bond between the carbonyl and the sulphur of the derivatized scaffold by adding a reagent that adds one or more metal chelating affinity ligand arms to the scaffold; and, if required, (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment of the method, steps (c) and (d) are performed simultaneously as one single step.

Abstract: The present invention relates to a method of generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of (a) providing at least one cyclic scaffold comprising a carbonyl, an adjacent sulphur and a nucleophile; (b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities are protected, on each scaffold by derivatisation of the nucleophile of said scaffolds, while retaining the cyclic structure of the scaffold; (c) ring-opening at the bond between the carbonyl and the sulphur of the derivatised scaffold by adding a reagent that adds one or more metal chelating affinity ligand arms to the scaffold; and, if required, (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment of the method, steps (c) and (d) are performed simultaneously as one single step.

Abstract: The present invention relates to an IgG-binding compound, which more specifically has affinity for human IgGs of ?-type and functional derivatives thereof. More specifically, the compound according to the invention comprises an N,N-alkylated urea moiety located between an aromatic part and another part, which is a linear or cyclic substituted or unsubstituted aliphatic group. The compound binds to a pocket-shaped binding site present on all human IgG ?-Fabs, which site is located between the two domains (CH1 and CL) of its constant part. Accordingly, the compound according to the invention is a ligand for human IgGs of ?-type, and consequently, the invention also relates to a separation matrix for affinity chromatography, which matrix comprises said compound, as well as to other uses of the compound.

Abstract: The present invention relates to a method of generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of