	US 11,680,081 B2
	Method for preparation of a separation matrix
Gustav Rodrigo, Uppsala (SE); Mats Ander, Uppsala (SE); Ronnie Palmgren, Uppsala (SE); and Tomas Bjorkman, Uppsala (SE)
Assigned to CYTIVA BIOPROCESS R&D AB, Uppsala (SE)
Appl. No. 16/955,119
Filed by Cytiva Bioprocess R&D AB, Uppsala (SE)
PCT Filed Dec. 13, 2018, PCT No. PCT/EP2018/084702 § 371(c)(1), (2) Date Jun. 18, 2020, PCT Pub. No. WO2019/121296, PCT Pub. Date Jun. 27, 2019.
Claims priority of application No. 1721476 (GB), filed on Dec. 20, 2017.
Prior Publication US 2021/0163529 A1, Jun. 3, 2021
Int. Cl. C07K 1/22 (2006.01); B01D 15/38 (2006.01); B01J 20/24 (2006.01); B01J 20/28 (2006.01); B01J 20/288 (2006.01); B01J 20/30 (2006.01); B01J 20/32 (2006.01); C07K 16/00 (2006.01)

CPC C07K 1/22 (2013.01) [B01D 15/3809 (2013.01); B01J 20/24 (2013.01); B01J 20/288 (2013.01); B01J 20/28016 (2013.01); B01J 20/3071 (2013.01); B01J 20/3212 (2013.01); B01J 20/3217 (2013.01); B01J 20/3274 (2013.01); B01J 20/3295 (2013.01); C07K 16/00 (2013.01); B01J 2220/54 (2013.01)]

48 Claims

1. A method for preparation of a separation matrix, comprising the steps of:

a) providing a solid support and an alkali-stable ligand derived from a bacterial immunoglobulin-binding protein;

b) reacting said alkali-stable ligand with said solid support to form a separation matrix having covalently coupled alkali-stable ligands; and

c) washing said separation matrix having covalently coupled alkali-stable ligands with a wash solution comprising at least 10 mM of an alkali metal hydroxide;

wherein the separation matrix is substantially free from non-covalently coupled ligands derived from said bacterial immunoglobulin-binding protein; and

wherein said alkali-stable ligand derived from a bacterial immunoglobulin-binding protein is capable of withstanding incubation with 0.1 M NaOH for 100×10 min at 22+/−2° C. with less than a 45% reduction in binding capacity towards IgG.