Synthetic single domain antibody

Patent metadata

Field	Value
Publication number	US-10030068-B2
Application number	US-201715638509-A
Country	US
Kind code	B2
Filing date	Jun 30, 2017
Priority date	Nov 4, 2013
Publication date	Jul 24, 2018
Grant date	Jul 24, 2018

Abstract

The invention relates to the identification of a highly stable single domain antibody scaffold (hs2d Ab) and its use in generating synthetic single domain antibody library (hs2d Ab-L1). The invention also relates to antigen-binding proteins comprising said stable single domain antibody scaffold and their uses, in particular as therapeutics.

First claim

The invention claimed is:

1. An antigen-binding protein, comprising a synthetic single domain antibody of the following formula: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, wherein said framework regions consist of FR1 of SEQ ID NO:1, FR2 of SEQ ID NO:2, FR3 of SEQ ID NO: 3 and FR4 of SEQ ID NO:4.

2. The antigen-binding protein of claim 1 , wherein said synthetic single domain antibody has one or more of the following functional properties:
i. it can be expressed as soluble single domain antibody in E. coli periplasm,
ii. it can be expressed as soluble intrabodies in E. coli cytosol,
iii. it is stable in reducing environment in chloramphenicol acetyl transferase fusion assay,
iv. it does not aggregate when expressed in mammalian cell lines as fluorescent protein fusions.

3. The antigen-binding protein of claim 1 , wherein the amino acid residues of the synthetic CDR1 and CDR2 are:
at CDR1 position 1: Y, R, S, T, F, G, A, or D;
at CDR1 position 2: Y, S, T, F, G, T, or T;
at CDR1 position 3: Y, S, F, or W;
at CDR1 position 4: Y, R, S, T, F, G, A, W, D, E, K or N;
at CDR1 position 5: S, T, F, G, A, W, D, E, N, I, H, R, Q, or L;
at CDR1 position 6: S, T, Y, D, or E;
at CDR1 position 7: S, T, G, A, D, E, N, I, or V;
at CDR2 position 1: R, S, F, G, A, W, D, E, or Y;
at CDR2 position 2: S, T, F, G, A, W, D, E, N, H, R, Q, L or Y;
at CDR2 position 3: S, T, F, G, A, W, D, E, N, H, Q, P;
at CDR2 position 4: G, S, T, N, or D;
at CDR2 position 5: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, V, W, K or M;
at CDR2 position 6: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, V, W, or K;
at CDR2 position 7: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, or V;
and wherein CDR3 amino acid sequence comprises between 9 and 18 amino acids selected among one or more of the following amino acids: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, V, W, K, M.

4. The antigen-binding protein of claim 1 , further comprising a F-box domain for targeting a protein to the proteasome.

Assignees

• Inst Nat Sante Rech Med
• Inst Curie
• Univ Toulouse 3 Paul Sabatier
• Centre Nat Rech Scient

Inventors

• Olichon Aurelien
• Moutel Sandrine
• Perez Franck

Classifications

• C07K16/44

  against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K16/32

  against translation products of oncogenes · CPC title

• C07K2317/569

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