Affinity carrier using mutant VHH antibody

Patent metadata

Field	Value
Publication number	US-12187764-B2
Application number	US-201917253372-A
Country	US
Kind code	B2
Filing date	Jun 20, 2019
Priority date	Jun 20, 2018
Publication date	Jan 7, 2025
Grant date	Jan 7, 2025

Abstract

Provision of an affinity carrier using a mutant VHH antibody. An affinity carrier comprising: a solid phase carrier; and an immunoglobulin-binding protein bound to the solid phase carrier; wherein the immunoglobulin-binding protein comprises a mutant VHH antibody or a fragment of the mutant VHH antibody that recognizes an epitope in at least one region selected from the group consisting of amino acids 127 to 184 of SEQ ID NO: 22 and amino acids 13 to 210 of SEQ ID NO: 23.

First claim

The invention claimed is:

1. An affinity carrier, comprising:
a solid phase carrier; and
an immunoglobulin-binding protein bound to the solid phase carrier,
wherein the immunoglobulin-binding protein comprises a mutant VHH antibody, or a fragment thereof, comprising a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO: 1, the polypeptide has the CDR1 and CDR2 amino acid sequences of SEQ ID NO: 10 and SEQ ID NO: 11, respectively, corresponding to amino acids 26 to 35 and 51 to 65 of SEQ ID NO: 1, the polypeptide has the CDR3 amino acid sequence of SEQ ID NO: 12 corresponding to amino acids 96 to 107 of SEQ ID NO: 1, in which at least at one position selected from the group consisting of amino acids 97, 100, and 102 of SEQ ID NO: 1 has been substituted with histidine, and the immunoglobulin-binding protein binds to an Fab domain of trastuzumab.

2. The affinity carrier according to claim 1 , wherein the immunoglobulin-binding protein comprises a polypeptide selected from the group consisting of SEQ ID NOS: 5, 6, 7 and 8.

3. An affinity carrier, comprising:
a solid phase carrier; and
an immunoglobulin-binding protein bound to the solid phase carrier,
wherein the immunoglobulin-binding protein comprises a mutant VHH antibody, or a fragment thereof, comprising a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO: 1, the polypeptide has at least one position selected from the group consisting of amino acids 43, 53, 64, 75, and 86 of SEQ ID NO: 1 substituted with arginine and retains the CDR1, CDR2, and CDR3 amino acid sequences of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, corresponding to amino acids 26 to 35, 51-65, and 96-107 of SEQ ID NO: 1 except for any substitution at amino acid positions 53 and 64 in CDR2, and the immunoglobulin-binding protein binds to an Fab domain of trastuzumab.

4. The affinity carrier according to claim 3 , wherein the immunoglobulin-binding protein comprises the polypeptide of SEQ ID NO: 9.

Assignees

• Jsr Corp

Inventors

• Hagihara Yoshihisa
• Akazawa Yoko
• Ito Yuji
• Matsuda Tomonari
• Kiyose Norihiko
• Miyazaki Nobuo
• Fukuta Tetsuo
• Mizuguchi Yusaku

Classifications

• C07K2317/569

  Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® · CPC title

• C07K17/00

  Carrier-bound or immobilised peptides (carrier-bound or immobilised enzymes C12N11/00); Preparation thereof · CPC title

• C07K16/4258

  against anti-receptor Ig · CPC title

• C07K16/00

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

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title