Fab REGION-BINDING PEPTIDE

Patent metadata

Field	Value
Publication number	US-2016289306-A1
Application number	US-201414914439-A
Country	US
Kind code	A1
Filing date	Aug 28, 2014
Priority date	Aug 30, 2013
Publication date	Oct 6, 2016
Grant date	—

Abstract

An object of the present invention is to provide a Fab region-binding peptide having an excellent ability for binding to a Fab region of IgG, an affinity separation matrix having the peptide as a ligand, and a method for producing a Fab region-containing protein, the method using the affinity separation matrix. Further, another object of the present invention is to provide a DNA encoding for the peptide, a vector containing the DNA, and a transformant which has been transformed by the vector. The Fab region-binding peptide according to the present invention is characterized in having a mutation at a specific site in comparison with wild-type SpG-β1.

First claim

1 . A Fab region-binding peptide represented by any one of the following (1) to (3):
(1) a Fab region-binding peptide having an amino acid sequence of SEQ ID NO: 1 derived from the β1 domain of protein G with substitution of an amino acid residue at not less than one position selected from the 13th, 19th, 30th and 33rd positions, and having a higher binding force to the Fab region of immunoglobulin than that before introduction of the substitution; (2) a Fab region-binding peptide having an amino acid sequence specified in the (1) with deletion, substitution and/or addition of not more than 1 and not more than 20 amino acid residues at a region other than the 13th, 19th, 30th and 33rd positions, and having a higher binding force to the Fab region of immunoglobulin than the peptide having the amino acid sequence of SEQ ID NO: 1; (3) a Fab region-binding peptide having an amino acid sequence having not less than 80% sequence identity to the amino acid sequence specified in the (1), and having a higher binding force to the Fab region of immunoglobulin than the peptide having the amino acid sequence of SEQ ID NO: 1, wherein the substituted amino acid residue at not less than 1 position selected from the 13th, 19th, 30th, and 33rd positions of the amino acid sequence specified in the (1) is not further mutated in the (3).

2 . The Fab region-binding peptide according to claim 1 , wherein the amino acid residue at the 13th position of the amino acid sequence specified in the (1) is substituted.

3 . The Fab region-binding peptide according to claim 2 , wherein the amino acid residue at the 13th position is substituted by Thr or Ser.

4 . The Fab region-binding peptide according to claim 1 , wherein the amino acid residue at the 30th position of the amino acid sequence specified in the (1) is substituted by Val, Leu or Ile.

5 . The Fab region-binding peptide according to claim 1 , wherein the amino acid residue at the 19th position of the amino acid sequence specified in the (1) is substituted by Val, Leu or Ile.

6 . The Fab region-binding peptide according to claim 1 , wherein the amino acid residue at the 33rd position of the amino acid sequence specified in the (1) is substituted by Phe.

7 . The Fab region-binding peptide according to claim 1 , wherein the position of deletion, substitution and/or addition of an amino acid residue is at least one position selected from the 2nd, 10th, 15th, 18th, 21st, 22nd, 23rd, 24th, 25th, 27th, 28th, 31st, 32nd, 35th, 36th, 39th, 40th, 42nd, 45th, 47th and 48th positions of the amino acid sequence specified in the (2).

8 . The Fab region-binding peptide according to claim 1 , wherein the position of deletion, substitution and/or addition of an amino acid residue is an N-terminus and/or a C-terminus of the amino acid sequence specified in the (2).

9 . The Fab region-binding peptide according to claim 1 , wherein the sequence identity to the amino acid sequence specified in the (3) is not less than 95%.

10 . A Fab region-binding peptide multimer, having not less than 2 domains wherein not less than 2 Fab region-binding peptides according to claim 1 are connected with one another.

11 . An affinity separation matrix, wherein a Fab region-binding peptide according to claim 1 or a Fab region-binding peptide multimer according to claim 10 is immobilized on a water-insoluble carrier as a ligand.

12 . A method for producing a Fab region-containing protein, comprising the steps of
contacting the affinity separation matrix according to claim 11 with a liquid sample containing the Fab region-containing protein, and separating the Fab region-containing protein bound to the affinity separation matrix from the affinity separation matrix.

13 . A DNA, encoding the Fab region-binding peptide according to claim 1 .

14 . A vector, containing the DNA according to claim 13 .

15 . A transformant, transformed by the vector according to claim 14 .

Assignees

• Kaneka Corp

Inventors

• Yoshida Shinichi
• Murata Dai

Classifications

• C12N2510/02

  Cells for production · CPC title

• C07K2317/55

  Fab or Fab' · CPC title

• C07K14/315

  from Streptococcus (G), e.g. Enterococci · CPC title

• C07K16/1275 Primary

  Streptococcus (G) · CPC title

• C07K1/22 Primary

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