US2017247407A1PendingUtilityA1

Affinity chromatography matrix

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Assignee: GE HEALTHCARE BIOPROCESS R&D ABPriority: Nov 30, 2011Filed: May 16, 2017Published: Aug 31, 2017
Est. expiryNov 30, 2031(~5.3 yrs left)· nominal 20-yr term from priority
Inventors:Hans J. JohanssonRonnie Palmgren
C07K 2319/00B01J 20/285B01J 20/3274B01D 15/34B01J 20/289C07K 1/22B01J 20/286B01D 15/3809C07K 14/31C07K 14/245
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Claims

Abstract

The invention discloses a polypeptide capable of binding immunoglobulins or immunoglobulin-containing proteins, which polypeptide comprises six or more domains of protein Z or the C domain of protein A or a functional variant thereof. It also discloses separation matrices comprising the polypeptide and methods of using the separation matrices for separation of immunoglobulins or immunoglobulin-containing proteins.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A separation matrix comprising a plurality of chromatography ligands coupled to a solid support,
 wherein the ligand comprises one or more mutated Domain C of Staphylococcus protein A (SpA) and having a sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO:1 or at least 77% identical to the amino acid sequence shown in SEQ ID NO:2.   
     
     
         2 . The separation matrix of  claim 1 , wherein the ligands after 5 hours incubation in 0.5 M NaOH has retained at least 95% of its original binding capacity. 
     
     
         3 . The separation matrix of  claim 1 , wherein the ligand is capable of binding to the Fab part of an antibody. 
     
     
         4 . The separation matrix of  claim 1 , wherein the ligand comprises one or more mutated Domain C of Staphylococcus protein A (SpA) and having a sequence that is at least 95% identical to the amino acid sequence shown in SEQ ID NO:1. 
     
     
         5 . The separation matrix of  claim 1 , wherein the ligand comprises one or more mutated Domain C of Staphylococcus protein A (SpA) and having a sequence that is at least 94% identical to the amino acid sequence shown in SEQ ID NO: 2. 
     
     
         6 . The separation matrix of  claim 1 , wherein the ligand comprises one or more mutated Domain C of Staphylococcus protein A (SpA) and having a sequence that is at least 98% identical to the amino acid sequence shown in SEQ ID NO:1 or SEQ ID NO: 2. 
     
     
         7 . The separation matrix of  claim 1 , wherein the one or more mutated Domain C of Staphylococcus protein A (SpA) comprises a sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 1 with mutation G29A, and SEQ ID NO: 2 with mutation G25A. 
     
     
         8 . The separation matrix of  claim 1 , wherein at least one asparagine in the sequence of the ligand is replaced with an amino acid other than glutamine or aspartic acid. 
     
     
         9 . The separation matrix of  claim 1 , wherein the ligand comprises a terminal coupling group comprising at least one nitrogen and/or sulphur atom(s). 
     
     
         10 . The separation matrix of  claim 9 , wherein the terminal group comprises arginine or cysteine. 
     
     
         11 . The separation matrix of  claim 1 , wherein the ligand is coupled to the solid support via thioether bonds. 
     
     
         12 . The separation matrix of  claim 1 , wherein the ligand is multimeric that contains at least two Domain C mutants. 
     
     
         13 . The separation matrix of  claim 12 , wherein the multimeric ligand comprises one or more other alkaline-stable protein-based units. 
     
     
         14 . The separation matrix of  claim 12 , wherein the multimeric ligand comprises 2-8 Domain C units, optionally coupled via linker segments. 
     
     
         15 . The separation matrix of  claim 1 , wherein the solid support is a polysaccharide. 
     
     
         16 . The separation matrix of  claim 1 , wherein the solid support is comprised of substantially spherical particles. 
     
     
         17 . The separation matrix of  claim 1 , wherein the solid support is porous. 
     
     
         18 . A separation matrix comprising a plurality of chromatography ligands coupled to a solid support,
 wherein the ligand comprises one or more mutated Domain C of Staphylococcus protein A (SpA) and having a sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 1 with mutation G29A, and SEQ ID NO: 2 with mutation G25A.   
     
     
         19 . The separation matrix of  claim 18 , wherein the ligand comprises a terminal coupling group and the ligand is coupled to the solid support via thioether bonds. 
     
     
         20 . The separation matrix of  claim 18 , wherein the solid support is porous spherical and comprises polysaccharide.

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