Immunoglobulin-binding protein and affinity carrier using same

Patent metadata

Field	Value
Publication number	US-10723769-B2
Application number	US-201615561332-A
Country	US
Kind code	B2
Filing date	Mar 24, 2016
Priority date	Mar 26, 2015
Publication date	Jul 28, 2020
Grant date	Jul 28, 2020

Abstract

Provided is an affinity chromatography carrier that maintains high immunoglobulin-binding capacity and high alkali resistance. An immunoglobulin-binding protein including at least one modified immunoglobulin-binding domain, the modified immunoglobulin-binding domain being a polypeptide consisting of an amino acid sequence of an immunoglobulin-binding domain selected from the group consisting of the B domain, Z domain, C domain, and variants thereof of Staphylococcus aureus protein A, in which at least one amino acid residue is inserted between positions corresponding to the 3-position and position 4 of the amino acid sequence of the B domain, Z domain or C domain.

First claim

The invention claimed is:

1. An immunoglobulin-binding protein comprising at least one modified immunoglobulin-binding domain,
wherein the at least one modified immunoglobulin-binding domain is selected from the group consisting of:
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 1, wherein at least one amino acid residue is inserted in said amino acid sequence between position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 1, wherein at least one amino acid residue is inserted in said amino acid sequence between positions corresponding to the position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 2, wherein at least one amino acid residue is inserted in said amino acid sequence between position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 2, wherein at least one amino acid residue is inserted in said amino acid sequence between positions corresponding to the position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 3, wherein at least one amino acid residue is inserted in said amino acid sequence between position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 3, and
a polypeptide consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 3, wherein at least one amino acid residue is inserted in said amino acid sequence between positions corresponding to the position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 3, and
wherein the at least one amino acid residue to be inserted is at least one selected from the group consisting of Ala, Arg, Asp, Gln, Glu, His, Met, Thr, Val, Phe, Leu, Ile, Pro, Trp, and Tyr.

2. The immunoglobulin-binding protein according to claim 1 , wherein the immunoglobulin-binding domain consisting of an amino acid sequence having at least 85% identity with an amino acid sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 3 is a Val1/Ala29 variant.

3. The immunoglobulin-binding protein according to claim 1 , comprising 2 to 12 units of the modified immunoglobulin-binding domain.

4. An affinity carrier comprising the immunoglobulin-binding protein according to claim 1 immobilized on a base material insoluble in water.

5. A method for isolating an immunoglobulin, the method comprising contacting a solution comprising an immunoglobulin with the affinity carrier according to claim 4 .

6. A method for producing an antibody medicine, the method comprising contacting a solution comprising an immunoglobulin with the affinity carrier according to claim 4 .

7. The immunoglobulin-binding protein according to claim 1 , wherein the at least one modified immunoglobulin-binding domain is selected from the group consisting of:
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 1, wherein at least one amino acid residue is inserted in said amino acid sequence between position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO: 1, wherein at least one amino acid residue is inserted in said amino acid sequence between positions corresponding to the position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 2, wherein at least one amino acid residue is inserted in said amino acid sequence between position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO: 2, wherein at least one amino acid residue is inserted in said amino acid sequence between positions corresponding to the position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 3, wherein at least one amino acid residue is inserted in said amino acid sequence between position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 3, and
a polypeptide consisting of an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO: 3, wherein at least one amino acid residue is inserted in said amino acid sequence between positions corresponding to the position 3 and position 4 of the amino acid sequence set forth in SEQ ID NO: 3, and
wherein the at least one amino acid residue to be inserted is at least one selected from the group consisting of Ala, Arg, Asp, Gln, Glu, His, Met, Thr, Val, Phe, Leu, Ile, Pro, Trp, and Tyr.

8. A method for producing an immunoglobulin-binding protein, the method comprising:
inserting, with regard to an amino acid sequence of an immunoglobulin-binding domain at least one amino acid residue between positions corresponding to the position 3 and position 4 of the amino acid sequence of the immunoglobulin-binding domain,
wherein the immunoglobulin-binding domain is selected from the group consisting of:
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 3, and
a polypeptide consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 3, and
wherein the at least one amino acid residue to be inserted is at least one selected from the group consisting of Ala, Arg, Asp, Gln, Glu, His, Met, Thr, Val, Phe, Leu, Ile, Pro, Trp, and Tyr.

9. The method according to claim 8 , wherein the immunoglobulin-binding domain consisting of an amino acid sequence having at least 85% identity with the amino acid sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 3 is a Val1/Ala29 variant.

10. The method according to claim 8 , further comprising linking 2 to 12 units of the immunoglobulin-binding domain having at least one amino acid residue inserted therein.

11. The method according to claim 8 , wherein the immunoglobulin-binding domain is selected from the group consisting of:
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO: 1,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO: 2,
a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 3, and
a polypeptide consisting of an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO: 3, and
wherein the at least one amino acid residue to be inserted is at least one selected fr

Assignees

• Jsr Corp
• Jsr Life Sciences Corp

Inventors

• Yasuoka Jun-Ichi
• Ichii Takashi
• Nakamura Satoshi
• Shiotani Tomonori
• Itaya Kaori

Classifications

• C07K17/04

  entrapped within the carrier, e.g. gel, hollow fibre · CPC title

• C07K16/065

  Purification, fragmentation · CPC title

• C07K14/31 Primary

  from Staphylococcus (G) · CPC title

• C07K1/22

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

• G01N33/6854

  Immunoglobulins · CPC title