Purification of immunoglobulins
View Patent ↗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.
1. A method of isolating antibodies from a liquid, which method comprises the steps of:
(a) providing a liquid that comprises at least one antibody;
(b) contacting said liquid with a separation matrix, which comprises one or more aliphatic sulphonamide ligands, to adsorb one or more antibodies to said matrix;
(c) passing an eluent over said matrix to release one or more antibodies; and
(d) recovering at least one antibody from a fraction of the eluent.
2. The method of claim 1 , wherein the liquid provided in step (a) additionally comprises one or more other proteins.
3. The method of claim 1 , wherein the separation matrix of step (b) is provided in a chromatography column.
4. The method of claim 3 , wherein step (b) is performed at a close to neutral pH.
5. The method of claim 1 , wherein the separation matrix of step (b) comprises a porous support; and ligands including one or more sulphonamides wherein an R group of the sulphonyl is an aliphatic compound; wherein said ligands are immobilized, optionally via spacer arms, on said porous support.
6. The method of claim 1 , wherein step (c) is a gradient elution performed by adding an eluent of decreasing salt concentration to the separation matrix.
7. The method of claim 1 , wherein step (b) is performed at a pH of or above neutral and step (c) is a gradient elution performed by adding an eluent of decreasing pH.
8. The method of claim 1 , wherein the antibodies recovered in step (d) are human or humanised antibodies.
9. The method of claim 1 , wherein the antibodies recovered in step (d) are immunoglobulin G (IgG).
10. The method of claim 1 , further comprising determining the amount of isolated antibody spectrophotometrically.