Ge Healthcare Bio-sciences Ab

Amersham Biosciences Ab

Pharmacia Biotech Ab

Amersham Pharmacia Biotech Ab

Bioresources International Inc.

Cytiva Bioprocess R&d Ab

Makonnen Belew

Jean-Luc Maloisel

Bo-Lennart Johansson

Jan Bergstrom

Wei Zhang

Lennart Soderberg

Rolf Berglund

Mei Yan Li

Mikael Andersson

Jan Gustavsson

Robert Asare Acquaah, Deceased

Daniel Gyingiri Achel

George Enyimah Armah

Jean-Luc Ge Healthcare Maloisel

A method for the removal of a substance, which has a negative charge and which is present in an aqueous liquid (I). The method comprises the steps of: (i) contacting the liquid with an anion-exchanger (1) that comprises mixed mode anion-exchanging ligands in which there is a positively charged nitrogen allowing binding of the substance to the anion-exchanger; and (ii) desorbing said substance from said anion-exchanger. The characteristic feature is that (A) the mixed mode ligands have a thioether linkage within a distance of 1-7 atoms from their positively charged atom, and (B) the anion-exchanger (1) (i) is capable of binding the substance of interest in an aqueous reference liquid (II) at an ionic strength corresponding to 0.25 M NaCl, and (ii) permits in the pH interval 2-12 a maximal breakthrough capacity for the substance which is &#x2267;200% of the breakthrough capacity of the substance for Q-Sepharose Fast Flow (anion-exchanger 2).

Method for anion-exchange adsorption and thioether anion-exchangers

The present invention is a method of purifying recombinant human serum albumin (rHSA) from a solution, which method comprises to subject a cell culture supernatant (CCS) comprising rHSA cation exchange on a bimodal high salt tolerant matrix; hydrophobic interaction chromatography (HIC); anion exchange; and recovering the purified rHSA. The bimodal high salt tolerant cation exchange matrix used enables performing a purification of a cell culture supernatant directly, in the sense that no further dilution thereof is necessary. Due to its high binding capacity, said bimodal cation exchange matrix also allows use of a smaller amount of matrix as compared to a corresponding conventional cation exchanger matrix. Accordingly, the present invention allows substantial savings as regards volumes and consequently operation costs.

Method for albumin purification

The present invention is a method of separating recombinant human serum albumin (rHSA) from low molecular weight contaminants in a liquid, which method comprises the steps of: (a) providing a separation medium, which includes anion-exchanging ligands coupled to a base matrix; and (b) contacting the liquid with the separation medium to adsorb the rHSA to the ligands. In one embodiment, the functional groups of the ligands are weak anion-exchanging groups, preferably secondary amines, and the density of ligands on the base matrix is relatively high.

Method, use and kit for separating albumin from contaminants in a liquid

The invention relates to a method for removing a positively charged substance from an aqueous liquid (I) by contacting the liquid with a cation-exchanger (1), possibly followed by a subsequent desorption of said substance. The cation-exchanger is selected to be capable of (a) binding to said substance by cation-exchange in an aqueous liquid reference (II) at an ionic strength corresponding to 0.3 M NaCl and (b) permitting a break through capacity for said substance200%, such as300% or500%, of the break-through capacity of said substance for a reference cation-exchanger (2) containing sulphopropyl groups &#x2014;CH2CH2CH2SO2O&#x2014;. The cation exchange ligands have an at least bimodal function by comprising a cation exchanging group and a separate hydrogen-bonding atom. The invention also relates to a method for testing the appropriateness of a cation-exchanger for removing a substance from a liquid and novel cation exchangers.

Adsorption method and ligands

Method for the removal of a substance from an aqueous liquid by ion exchange comprising providing a liquid with said substance is present; providing an adsorption matrix which comprises at least two different ligands; contacting the liquid with the matrix under a period of time and conditions sufficient to allow adsorption of the substance to the matrix; and adding an eluent that desorbs the substance from the matrix. Each one of the ligands interacts with the substance during the adsorption step, and at least one of the ligands is charged and capable of ionic interaction with the substance. The method can be run as a cation or anion exchange. The substance desorption can be performed by adding an eluent comprising an increasing ionic strength. The invention also encompasses an adsorbent comprising at least two structurally different ligands, which interact with the same kind of substance when used in a separation procedure.

Method for mixed mode adsorption

Method for desalting an aqueous liquid containing a charged substance comprising the steps of: (i) contacting liquid (I) with an ion-exchanger (1) under conditions permitting binding between ion-exchanger and substance, said ion-exchanger comprising a base matrix carrying an ion-exchange ligand (Ligand 1) with the opposite charge compared to the substance, (ii) desorbing said substance from said ion-exchanger by the use of a liquid (liquid (II)). Wherein: (A) ion-exchanger (1) is an ion-exchanger: (a) that can bind the substance in an aqueous reference liquid at ionic strength corresponding to 0.1 M NaCl, preferably 0.25 M NaCl; and (b) permits a breakthrough capacity at the pH provided by liquid (I) which is more than 2 mg/ml of gel, at a breakthrough of 10% and linear flow velocity of 300 cm/h; and (B) in step (ii) the pH of liquid (11) is adjusted to a pH value where the charge difference between the substance, the ligand and/or the ion-exchanger is lowered.

Method for the manufacture of compositions containing low concentrations of salts

A method for the removal of a substance carrying a negative charge and being present in an aqueous liquid (I). The method comprises the steps of: (i) contacting the liquid with a matrix carrying a plurality of ligands comprising a positively charged structure and a hydrophobic structure, and (ii) desorbing the substance. The characterizing feature is that (I) each of said ligands together with a spacer has the formula: &mdash;SP&mdash;&lsqb;Ar&mdash;R &mdash;N (R R R )&rsqb; where (A) &lsqb;Ar&mdash;R &mdash;N (R R R )&rsqb; represents a ligand a) Ar is an aromatic ring, b) R is &lsqb;(L) R&prime; &rsqb; where n and m are integers selected amongst zero or 1; L is amino nitrogen, ether oxygen or thioether sulphur; R&prime; is a linker selected among 1) hydrocarbon groups; 2) &mdash;C(&boxH;NH)&mdash;; c) R are selected among hydrogen and alkyls; (B) SP is a spacer providing a carbon or a heteroatom directly attached to Ar&mdash;R &mdash;N (R R R ); (C)&mdash;represents that SP replaces a hydrogen in (Ar&mdash;R &mdash;N (R R R ); (D)&mdash;represents binding to the matrix; and (II) desorption. There is also described (a) anion-exchangers having high breakthrough capacities, (b) a screening method and (c) a desalting protocol.

Method for anion-exchange adsorption and anion-exchangers

Process for chromatographic separation of peptides and nucleic acid, and

A method for purifying a plant protein comprises: (a) preparing a crude plant extract; (b) passing the crude plant extract with a first buffer solution through a guard column comprising a hydrophobic interaction chromatography medium of hydrophobicity sufficiently high to prevent tannins and polyphenols from eluting from the column in the presence of the buffer solution, but sufficiently low that a protein fraction elutes with the first buffer solution; (c) passing the protein fraction through a capture column coupled in series to the guard column, the capture column comprising a hydrophobic interaction chromatography medium of hydrophobicity sufficiently high to prevent the protein from eluting in the presence of the buffer solution; and (d) eluting the protein from the capture column as a purified fraction. Preferably the plant is miracle fruit, the protein is Miraculin, and the method comprises ion exchange chromatography and gel filtration chromatography of the purified fraction of the protein. Foods and beverages may be sweetened with the purified Miraculin.

Growing community of inventors

Uppsala, Sweden

Makonnen Belew