Modified kappa light chain-binding polypeptides

Patent metadata

Field	Value
Publication number	US-11136357-B2
Application number	US-201916252015-A
Country	US
Kind code	B2
Filing date	Jan 18, 2019
Priority date	Dec 17, 2014
Publication date	Oct 5, 2021
Grant date	Oct 5, 2021

Abstract

A kappa light chain-binding polypeptide comprising or consisting essentially of one or more mutated binding domains of Peptostreptococcus Protein L.

First claim

The invention claimed is:

1. A kappa light chain-binding polypeptide comprising a mutated domain 3 of Peptostreptococcus Protein L and having an amino acid sequence specified by SEQ ID NO:4, wherein at least the amino acid at the position corresponding to position 45 has been mutated to alanine or the position corresponding to position 19 has been mutated to glutamic acid; or wherein the amino acid at the position corresponding to position 45 has been mutated to alanine or the position corresponding to position 19 has been mutated to glutamic acid with further mutations in at least one of the amino acid at the position corresponding to position 10 has been mutated to glutamine and the amino acid at the position corresponding to position 60 has been mutated to glutamine; and wherein the polypeptide binds the kappa light chain of an antibody or fragment thereof and has improved alkaline stability compared to a non-mutated domain 3 of Peptostreptococcus Protein L.

2. The polypeptide of claim 1 , comprising a multimer of mutated domain 3.

3. The polypeptide of claim 1 , wherein the amino acid at the position corresponding to position 10 has been mutated to glutamine.

4. The polypeptide of claim 1 , wherein the amino acid at the position corresponding to position 10 and 60 has been mutated to glutamine.

5. The polypeptide of claim 1 , wherein the amino acid at the position corresponding to position 60 has been mutated to glutamine.

6. The polypeptide of claim 1 , wherein the amino acid at the position corresponding to position 19 in has been mutated to glutamic acid.

7. The polypeptide of claim 1 , wherein the amino acid sequence of mutated domain 3 is selected from the group consisting of sequences defined by SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:11.

8. The polypeptide of claim 1 , wherein the amino acid sequence of mutated domain 3 is the amino acid sequence of SEQ ID NO:11.

9. The polypeptide of claim 1 , further comprising two, three, four, five, six or seven such mutated domain 3 amino acid sequences.

10. The polypeptide of claim 9 , wherein the domains are linked to each other by elements comprising up to five or fifteen amino acid residues.

11. The polypeptide of claim 1 , wherein the amino acid sequence of mutated domain 3 is SEQ ID NO:14.

12. A nucleic acid or a vector encoding a polypeptide according to claim 1 .

13. An expression system, which comprises a nucleic acid or vector according to claim 12 .

14. A separation matrix, wherein a plurality of polypeptides of claim 1 have been coupled to a solid support.

15. The separation matrix according to claim 14 , wherein the binding capacity of the matrix for kappa light chain-containing proteins after 100 15 min incubation cycles in 0.1 M NaOH at 22+/−2° C. is at least 40% of the binding capacity before the incubation.

Assignees

• Cytiva Bioprocess R & D Ab

Inventors

• Rodrigo Gustav
• Ander Mats
• Bjorkman Tomas

Classifications

• C07K2319/02

  containing a signal sequence · CPC title

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• B01D15/3809

  of the antigen-antibody type, e.g. protein A, G or L chromatography · CPC title

• C07K14/195 Primary

  from bacteria · CPC title

• B01J20/24

  Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives · CPC title