	US 11,753,438 B2
	Method of cleaning and/or sanitizing a separation matrix
Annika Forss, Uppsala (SE); Gustav José Rodrigo, Uppsala (SE); Tomas Bjorkman, Uppsala (SE); Mats Ander, Uppsala (SE); and Jesper Ulf Hansson, Uppsala (SE)
Assigned to CYTIVA BIOPROCESS R&D AB, Uppsala (SE)
Appl. No. 16/95,753
Filed by CYTIVA BIOPROCESS R&D AB, Uppsala (SE)
PCT Filed May 10, 2017, PCT No. PCT/EP2017/061159 § 371(c)(1), (2) Date Oct. 23, 2018, PCT Pub. No. WO2017/194593, PCT Pub. Date Nov. 16, 2017.
Application 16/095,753 is a continuation of application No. 15/348,699, filed on Nov. 10, 2016, granted, now 10,703,774.
Application 15/348,699 is a continuation of application No. 15/282,367, filed on Sep. 30, 2016, granted, now 10,654,887.
Application 15/348,699 is a continuation in part of application No. 15/282,367, filed on Sep. 30, 2016, granted, now 10,654,887.
Claims priority of application No. 1608229 (GB), filed on May 11, 2016; and application No. 1608232 (GB), filed on May 11, 2016.
Prior Publication US 2020/0299325 A1, Sep. 24, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 1/22 (2006.01); B01J 20/26 (2006.01); B01J 20/285 (2006.01); B01J 20/286 (2006.01); B01J 20/32 (2006.01); C07K 14/31 (2006.01); C07K 16/00 (2006.01); C07K 16/12 (2006.01); C07K 17/10 (2006.01)

CPC C07K 1/22 (2013.01) [B01J 20/267 (2013.01); B01J 20/285 (2013.01); B01J 20/286 (2013.01); B01J 20/3274 (2013.01); C07K 14/31 (2013.01); C07K 16/00 (2013.01); C07K 16/1271 (2013.01); C07K 17/10 (2013.01); B01J 2220/52 (2013.01)]

18 Claims

1. A method of cleaning and/or sanitizing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support, wherein the method comprises the steps of:

a) optionally purifying a mixture comprising a first immunoglobulin using the separation matrix;

b) providing a cleaning liquid comprising at least 50% by volume of an aqueous alkali metal hydroxide solution; and

c) cleaning and/or sanitizing the separation matrix by contacting the cleaning liquid with the separation matrix for a predetermined contact time;

wherein the alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA), wherein the parental Fc-binding domain of SpA is defined by SEQ ID NO: 51 or SEQ ID NO: 52 in that the amino acid residues at positions 13 and 44 of SEQ ID NO: 51 or 52 are asparagines, and wherein at least the asparagine residue at position 3, except the asparagine residues at positions 13 and 14, of SEQ ID NO: 51 or 52 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.