Immunoglobulin-binding protein, and affinity carrier using same

Patent metadata

Field	Value
Publication number	US-12428451-B2
Application number	US-201917268804-A
Country	US
Kind code	B2
Filing date	Aug 23, 2019
Priority date	Aug 24, 2018
Publication date	Sep 30, 2025
Grant date	Sep 30, 2025

Abstract

An affinity carrier may have improved alkali resistance. An immunoglobulin-binding protein containing a mutant polypeptide chain, as well as an affinity carrier containing a solid-phase carrier to which the immunoglobulin-binding protein is bound, may be a mutant polypeptide chain having an amino acid sequence having at least 85% identity to an amino acid sequence indicated by any of SEQ ID NO: 1-6 and 57-62, having a predetermined mutation, and having immunoglobulin-binding activity.

First claim

The invention claimed is:

1. An immunoglobulin-binding protein, comprising:
a polypeptide chain consisting of an amino acid sequence having at least 85% identity to an amino acid sequence represented by any one of SEQ ID NOs: 1 to 6, 57-59 and 61-62, wherein said amino acid sequence having at least 85% identity to any one of SEQ ID NOS: 1 to 6, 57-59 and 61-62 having a substitution of Asn with Gln at a position corresponding to position 11 of the amino acid sequence represented by SEQ ID NO: 1, 2, 3, 5, or at position 6 of the amino acid sequence represented by SEQ ID No. 4, or at position 9 of the amino acid sequence represented by SEQ ID No. 6, or at position 7 of the amino acid sequence represented by SEQ ID No. 57, 58, 59, 61, 62 and optionally having at least one mutation selected from the group consisting of (a) to (f):
(a) substitution of an amino acid residue at at least one position selected from the group consisting of positions corresponding to positions 3 and 6 of the amino acid sequence represented by SEQ ID NO: 3 with another amino acid residue, deletion of the amino acid residue at the at least one position, or insertion of another amino acid residue into a position in front of or behind the at least one position;
(b) substitution of an amino acid residue at at least one position selected from the group consisting of positions corresponding to positions 24 and 25 of the amino acid sequence represented by SEQ ID NO: 3 with another amino acid residue, deletion of the amino acid residue at the at least one position, or insertion of another amino acid residue into a position in front of or behind the at least one position;
(c) substitution of an amino acid residue at a position corresponding to position 39 of the amino acid sequence represented by SEQ ID NO: 3 with another amino acid residue, deletion of the amino acid residue at the position corresponding to position 39, or insertion of another amino acid residue into a position in front of or behind the position corresponding to position 39;
(d) substitution of an amino acid residue at a position corresponding to position 46 of the amino acid sequence represented by SEQ ID NO: 3 with another amino acid residue, deletion of the amino acid residue at the position corresponding to position 46, or insertion of another amino acid residue into a position in front of or behind the position corresponding to position 46;
(e) substitution of an amino acid residue at at least one position selected from the group consisting of positions corresponding to positions 4, 49, and 58 of the amino acid sequence represented by SEQ ID NO: 3 with another amino acid residue, deletion of the amino acid residue at the at least one position, or insertion of another amino acid residue into a position in front of or behind the at least one position; and
(f) substitution of an amino acid residue at a position corresponding to position 23 of the amino acid sequence represented by SEQ ID NO: 3 with another amino acid residue, deletion of the amino acid residue at the position corresponding to position 23, or insertion of another amino acid residue into a position in front of or behind the position corresponding to position 23.

2. The protein of claim 1 , wherein the polypeptide chain is a polypeptide chain consisting of an amino acid sequence having at least 85% identity to the amino acid sequence represented by SEQ ID NO: 3 and having the at least one mutation.

3. The protein of claim 1 , wherein the at least one mutation is selected from the group consisting of:
(a1) substitution of Asn with Gln at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a2) substitution of Asn with Ala at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a3) substitution of Asn with Asp at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a4) substitution of Asn with Gln at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a5) substitution of Asn with Ala at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a6) substitution of Asn with Asp at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(b1) substitution of Glu with Asp at a position corresponding to position 24 of the amino acid sequence represented by SEQ ID NO: 3;
(b2) substitution of Glu with Asp at a position corresponding to position 25 of the amino acid sequence represented by SEQ ID NO: 3;
(c1) substitution of Ser with Lys at a position corresponding to position 39 of the amino acid sequence represented by SEQ ID NO: 3;
(c2) substitution of Ser with Arg at a position corresponding to position 39 of the amino acid sequence represented by SEQ ID NO: 3;
(d1) substitution of Ala with Asp at a position corresponding to position 46 of the amino acid sequence represented by SEQ ID NO: 3;
(d2) substitution of Ala with Glu at a position corresponding to position 46 of the amino acid sequence represented by SEQ ID NO: 3;
(d3) substitution of Ala with Lys at a position corresponding to position 46 of the amino acid sequence represented by SEQ ID NO: 3;
(d4) substitution of Ala with Arg at a position corresponding to position 46 of the amino acid sequence represented by SEQ ID NO: 3;
(e1) deletion of Lys at a position corresponding to position 4 of the amino acid sequence represented by SEQ ID NO: 3;
(e2) substitution of Lys with Arg at a position corresponding to position 49 of the amino acid sequence represented by SEQ ID NO: 3;
(e3) deletion of Lys at a position corresponding to position 58 of the amino acid sequence represented by SEQ ID NO: 3;
(e4) substitution of Lys with Arg at a position corresponding to position 58 of the amino acid sequence represented by SEQ ID NO: 3;
(f1) substitution of Thr with Arg at a position corresponding to position 23 of the amino acid sequence represented by SEQ ID NO: 3;
(f2) substitution of Thr with Leu at a position corresponding to position 23 of the amino acid sequence represented by SEQ ID NO: 3; and
(f3) substitution of Thr with Ser at a position corresponding to position 23 of the amino acid sequence represented by SEQ ID NO: 3.

4. The protein of claim 3 , wherein the at least one mutation is selected from the group consisting of:
(a1) substitution of Asn with Gln at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a2) substitution of Asn with Ala at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a3) substitution of Asn with Asp at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a4) substitution of Asn with Gln at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a5) substitution of Asn with Ala at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a6) substitution of Asn with Asp at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a23) substitution of Asns with Glns at positions corresponding to positions 3 and 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a24) substitution of Asn with Ala at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a25) substitution of Asn with Asp at a position corresponding to position 3 of the amino acid sequence represented by SEQ ID NO: 3;
(a26) substitution of Asn with Asp at a position corresponding to position 6 of the amino acid sequence represented by SEQ ID NO: 3;
(a27) substitution of Asn with Asp at a position corresponding to position 3, and

Assignees

• Jsr Corp
• Jsr Life Sciences Llc
• Jsr Micro N V

Inventors

• Haga Tomoaki
• Ichii Takashi
• Onogi Shunsuke
• Nakamura Satoshi
• Honda Toshinari
• Kamide Tomoyuki

Classifications

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• C07K1/22

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

• B01D15/3809 Primary

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

• B01J20/3274

  Proteins, nucleic acids, polysaccharides, antibodies or antigens · CPC title

• C07K14/31 Primary

  from Staphylococcus (G) · CPC title