# US 9,040,666 B2 · Single-domain antigen-binding proteins that bind mammalian IgG

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- Number: US 9,040,666 B2
- Document ID: US9040666B2
- Status: Granted
- Kind: B2
- Application number: 12/669,034
- Filed: 2008-07-08
- Published: 2010-12-09
- Granted: 2015-05-26
- Earliest claimed: 2007-07-13
- Assignees: Bac IP B.V.
- Applicants: Wilhelmus Josephus Johanna Hermans; Sandra Bezemer; Yvonne Mathalie Mijnsbergen
- Inventors: Wilhelmus Josephus Johanna Hermans; Sandra Bezemer; Yvonne Mathalie Mijnsbergen
- Web page: https://patentgrep.com/patent/US9040666B2

## CPC

- C07K 16/4283: {against an allotypic or isotypic determinant on Ig}
- A61K 47/6843: {the antibody targeting a material from animals or humans}
- A61P 37/00: Drugs for immunological or allergic disorders
- C07K 16/18: against material from animals or humans
- C07K 2317/22: from camelids, e.g. camel, llama or dromedary
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/569: Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present application relates to antigen-binding proteins that are capable of binding to mammalian IgG. The frame-work regions of the antigen-binding proteins of the application preferably correspond to those of antibodies naturally that are devoid of light chains as may e.g. be found in camelids. The application further relates to nucleic acids that encode such antigen-binding proteins, to immunoadsorbent materials that comprise such proteins, to the uses of such immunoadsorbent materials for the purification of mammalian IgG antibodies and for therapeutic apheresis.

## Claims

### Claim 1

An antigen-binding protein comprising an amino acid sequence that comprises 4 framework regions, FR1 to FR4, and 3 complementarity determining regions, CDR1 to CDR3, that are operably linked in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, wherein:
(a) the CDR1 has an amino acid sequence as set forth in SEQ ID NO: 1;
(b) the CDR2 has an amino acid sequence as set forth in SEQ ID NO: 50; and,
(c) the CDR3 is an amino acid sequence as set forth in SEQ ID NO: 99; and,
wherein the antigen-binding protein has an amino acid sequence as set forth in SEQ ID NO 148, and wherein the antigen-binding protein specifically binds to the Fc domain of a human IgG molecule.

### Claim 2 (depends on 1)

The antigen-binding protein according to claim 1, wherein the antigen binding protein has one or more properties selected from the group consisting of:
(a) the antigen-binding protein binds the human IgG molecule with a binding affinity of at least 10-7 M as analyzed by surface plasmon resonance analysis (SPR) using polyclonal human IgG;
(b) the antigen-binding protein has a dynamic binding capacity of at least 2 mg human IgG/ml resin, when coupled to N-hydroxysuccinimide (NHS) activated carrier at a density of 20 mg antigen-binding protein per ml NHS resin and using a flow-rate of 150 cm/h.

### Claim 3 (depends on 1)

The antigen-binding protein according to claim 1, wherein the human IgG is an IgG1, IgG2, IgG3 or IgG4 molecule.

### Claim 4 (depends on 1)

A multivalent antigen-binding protein comprising the amino acid sequences of at least two antigen-binding proteins according to claim 1.

### Claim 5 (depends on 1)

The antigen-binding protein according to claim 1, wherein the antigen-binding protein is part of a fusion protein further comprising an amino acid sequence of a therapeutic protein or peptide.

### Claim 6

An antigen-binding protein comprising an amino acid sequence that comprises 4 framework regions, FR1 to FR4, and 3 complementarity determining regions, CDR1 to CDR3, that are operably linked in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, wherein:
(i) the CDR1 has an amino acid sequence as set forth in SEQ ID NO: 1;
(ii) the CDR2 has an amino acid sequence as set forth in SEQ ID NO: 50; and,
(iii) the CDR3 is an amino acid sequence as set forth in SEQ ID NO: 99; and,
wherein: FR1 comprises an amino acid selected from the group consisting of:
(a) an amino acid having SEQ ID: 197;
(b) an amino acid having at least 90% sequence identity with SEQ ID: 197; and/or,
(c) the amino acid sequence of a) that has one or more amino acid substitutions as defined in Table 1;
FR2 comprises an amino acid selected from the group consisting of:
(d) an amino acid having SEQ ID: 198;
(e) an amino acid having at least 90% sequence identity with SEQ ID: 198; and/or,
(f) the amino acid sequence of d) that has one or more amino acid substitutions as defined in Table 2;
FR3 comprises an amino acid selected from the group consisting of the amino acid sequence:
(g) an amino acid having SEQ ID: 199;
(h) an amino acid having at least 90% sequence identity with SEQ ID: 199; and/or,
(i) the amino acid sequence of g) that has one or more amino acid substitutions as defined in Table 3; and,
FR4 comprises an amino acid selected from the group consisting of:
(j) an amino acid having SEQ ID: 200;
(k) an amino acid having at least 90% sequence identity with SEQ ID: 200; and/or,
l) the amino acid sequence of j) that has one or more amino acid substitutions as defined in Table 4, and wherein the antigen-binding protein specifically binds to the Fc domain of a human IgG molecule.

### Claim 7 (depends on 6)

The antigen-binding protein according to claim 6, wherein the framework has at least 90% sequence identity with an amino acid sequence as set forth in SEQ ID NO:148.

### Claim 8 (depends on 6)

The antigen-binding protein according to claim 6, wherein the framework has at least 95% sequence identity with an amino acid sequence as set forth in SEQ ID NO: 148.