IP Library › Granted Patent US 12,337,293
Granted Patent B2
US 12,337,293 · App. 17/981,262 · Granted Jun 24, 2025

Chromatography ligand comprising domain c from

Inventors: Martin Hall (Uppsala, SE); Sture Larsson (Uppsala, SE); Andreas Muranyi (Uppsala, SE); Gustav Rodrigo (Uppsala, SE); Jinyu Zou (Uppsala, SE); Per-Mikael Aberg (Uppsala, SE)
Assignee: Cytiva Bioprocess R&D AB
B01J20/24B01D15/3809B01J20/28019B01J20/285B01J20/286B01J20/289B01J20/3212B01J20/3219B01J20/3274B01J20/3293C07K1/22C07K14/31C07K16/00C07K17/00C07K17/10B01J2220/52B01J2220/54C07K2317/55
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Quick Facts
Patent No.
US 12,337,293
App. No.
17/981,262
Granted
Jun 24, 2025
Kind
B2
Abstract

The present invention relates to a chromatography ligand, which comprises Domain C from Staphylococcus protein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIP) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates to a process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.

Claims (16)

1. A chromatography ligand, which ligand comprises one or more Domain C units from Staphylococcus protein A (SpA), wherein the Domain C sequence has a sequence that is at least 94% identical to an amino acid sequence, the amino acid sequence being a sequence shown in SEQ ID NO:2 with mutation G25A.

2. The chromatography ligand according to claim 1 , wherein after 15 hours incubation in 0.5 M NaOH the ligand retains at least 80% of its binding capacity compared to the untreated ligand.

3. The chromatography ligand according to claim 1 , wherein after 15 hours incubation in 0.5 M NaOH the ligand retains at least 90% of its binding capacity compared to the untreated ligand.

4. The chromatography ligand according to 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.

5. The chromatography ligand according to claim 1 , wherein the ligand is multimeric and comprises at least two Domain C mutants.

6. The chromatography ligand according to claim 1 , wherein the multimeric ligand comprises 2-8 Domain C units, optionally coupled via linker segments.

7. The chromatography ligand according to claim 1 , wherein the ligand comprises a terminal coupling group comprising at least one nitrogen and/or sulfur atom(s).

8. The chromatography ligand according to claim 7 , wherein the terminal group comprises arginine or cysteine.

9. A separation matrix comprising the ligand of claim 1 .

10. The separation matrix according to claim 9 , wherein after 15 hours incubation in 0.5 M NaOH the ligand retains at least 80% of its binding capacity compared to the untreated ligand.

11. The separation matrix according to claim 9 , wherein after 15 hours incubation in 0.5 M NaOH the ligand retains at least 90% of its binding capacity compared to the untreated ligand.

12. The separation matrix according to claim 9 , wherein at least one asparagine in the sequence of the ligand is replaced with an amino acid other than glutamine or aspartic acid.

13. The separation matrix according to claim 9 , wherein the ligand is multimeric and comprises at least two Domain C mutants.

14. The separation matrix according to claim 9 , wherein the multimeric ligand comprises 2-8 Domain C units, optionally coupled via linker segments.

15. The separation matrix according to claim 9 , wherein the ligand comprises a terminal coupling group comprising at least one nitrogen and/or sulfur atom(s).

16. The separation matrix according to claim 15 , wherein the terminal group comprises arginine or cysteine.

Priority Claims (1)
SE 0602061-4 · Sep 29, 2006 · national
Continuity (9)
Continuation 17107600 · Nov 30, 2020
Continuation 16443600 · Jun 17, 2019
Continuation 16189894 · Nov 13, 2018
Continuation 15603285 · May 23, 2017
Continuation 15063471 · Mar 7, 2016
Division 14164519 · Jan 27, 2014
Continuation In Part 13559663 · Jul 27, 2012
Division 12443011
Related Publication 20230347316A1 · Nov 2, 2023
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