Purification of immunoglobulins

Abstract

The present invention relates to a separation matrix comprised of a porous support to which ligands have been immobilised, 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 separation matrix comprising 
 (a) a porous support; and    (b) 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.    
   
   
       2 . The matrix of  claim 1 , wherein the sulphonamide is coupled to the porous support via its nitrogen.  
   
   
       3 . The matrix of  claim 1 , wherein the sulphonamide is coupled to the porous support via its sulphur.  
   
   
       4 . The matrix of  claim 1 , wherein the R group is a methyl group.  
   
   
       5 . The matrix of  claim 1 , wherein the nitrogen of the sulphonamide(s) is a primary or secondary amine.  
   
   
       6 . The matrix of  claim 1 , wherein the ligands are monoamines.  
   
   
       7 . The matrix of  claim 1 , wherein the ligands are polyamines.  
   
   
       8 . The matrix of  claim 7 , wherein each polyamine comprises two to six amines.  
   
   
       9 . The matrix of  claim 1 , wherein the ligands are present as repetitive units of a polymer immobilised to the support.  
   
   
       10 . The matrix of  claim 9 , wherein the polymer is a polyethylene imine.  
   
   
       11 . The matrix of  claim 9 , wherein the polymer exhibit two or more different ligand groups.  
   
   
       12 . The matrix of  claim 1 , wherein the ligands are aliphatic compounds.  
   
   
       13 . The matrix of  claim 1 , wherein the support is a cross-linked polysaccharide.  
   
   
       14 . A chromatography column packed with the separation matrix of  claim 1 .  
   
   
       15 . The chromatography column of  claim 14 , which is substantially sterile.  
   
   
       16 . The chromatography column of  claim 14 , which is a disposable column.  
   
   
       17 . A process of preparing a matrix for separation of antibodies, which method comprises a first step of immobilising amines and/or polyamines to a porous support and a subsequent step of sulphonylating said amines to provide aliphatic sulphonamide ligands.  
   
   
       18 . A process of preparing a matrix for separation of antibodies, which method comprises a first step of activating a porous support and a subsequent step of attaching sulphonamides to the activated sites via their sulphurs to provide aliphatic sulphonamide ligands.  
   
   
       19 . 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; and, optionally,    (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.    
   
   
       20 . The method of  claim 19 , wherein the liquid provided in step (a) additionally comprises one or more other proteins.  
   
   
       21 . The method of  claim 19 , wherein the separation matrix of step (b) is provided in a chromatography column.  
   
   
       22 . The method of  claim 19 , wherein the separation matrix of step (b) is as defined in  claim 1 .  
   
   
       23 . The method of  claim 21 , wherein step (b) is performed at a close to neutral pH.  
   
   
       24 . The method of  claim 19 , wherein step (c) is a gradient elution performed by adding an eluent of decreasing salt concentration to the separation matrix.  
   
   
       25 . The method of  claim 19 , 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.  
   
   
       26 . The method of  claim 19 , wherein the antibodies recovered in step (d) are human or humanised antibodies.  
   
   
       27 . The method of  claim 19 , wherein the antibodies recovered in step (d) are immunoglobulin G (IgG).  
   
   
       28 . The method of  claim 19 , further comprising determining the amount of isolated antibody spectrophotometrically.