Granted Patent B2

US 12,221,492 · App. 17/149,830 · Granted Feb 11, 2025

Inventors: Gustav Rodrigo (Uppsala, SE); Tomas Bjorkman (Uppsala, SE); Mats Ander (Uppsala, SE)

Assignee: Cytiva BioProcess R&D AB

C07K16/46B01D15/3809B01J20/289B01J20/3219B01J20/3274C07K1/22C07K14/31C07K16/00C07K16/065

Abstract

Claims (22)

1. An Fc-binding polypeptide comprising a mutant of an Fc-binding domain of Staphylococcus Protein A (SpA) as defined by a consensus sequence KEX 1 QX 2 AFYEILX 3 LPNLTEEQRNAFIQX 6 LKDX 7 PSQSX 9 X 10 X 11 LAEAKX 12 X 13 ND AQAPK (SEQ ID NO:28), wherein X 7 is E and X 9 is R, and wherein individually of each other at least one of X 1 , X 2 , X 3 , X 6 , X 10 , X 11 , X 12 , and X 13 is substituted as follows:

X 1 =A or else Q,

X 2 =K or else or N,

X 3 =K or else H,

X 6 =K or else S,

X 10 =A or else N,

X 11 =I or else L,

X 12 =R or else K,

X 13 =Y or else L,

and wherein the mutant binds the Fc domain of an antibody and has improved alkaline stability compared to the amino acid sequence of SEQ ID NO:7, wherein the alkaline stability is improved as measured by the remaining IgG-binding capacity after 24 or 25 h incubation in 0.5 M or 1.0 M aqueous NaOH at 22+/−2° C.

2. The polypeptide of claim 1 , wherein X 1 =A.

3. The polypeptide of claim 1 , wherein X 2 =K.

4. The polypeptide of claim 1 , wherein X 3 =K.

5. The polypeptide of claim 1 , wherein X 10 =N.

6. The polypeptide of claim 1 , wherein X 11 =L.

7. The polypeptide of claim 1 , wherein X 12 =K.

8. The polypeptide of claim 1 , wherein X 13 =Y.

9. The polypeptide of claim 1 , wherein the mutant is selected from the group consisting of SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO:11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, and SEQ ID NO:17.

10. A multimer comprising or consisting essentially of a plurality of polypeptides as defined by claim 1 .

11. The multimer of claim 10 , wherein the polypeptides are linked by linkers comprising up to 15 amino acids.

12. The multimer of claim 10 , which is a dimer, trimer, tetramer, pentamer, hexamer, heptamer, octamer or nonamer.

13. The multimer of claim 10 , which comprises a sequence as defined by SEQ ID NO: 20, SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23.

Assignments (3)

Priority Claims (3)

Continuity (2)

References Cited (300)