# US 7,608,695 B2 · Substance with antithrombotic activity and method for detecting glycokallidin

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- Number: US 7,608,695 B2
- Document ID: US7608695B2
- Status: Granted
- Kind: B2
- Application number: 10/825,127
- Filed: 2004-04-16
- Published: 2004-11-18
- Granted: 2009-10-27
- Earliest claimed: 1998-04-23
- Assignees: Ajinomoto Co., Inc.
- Inventors: Naoyuki Fukuchi; Fumie Futaki; Morikazu Kito; Seiichi Sato; Takayuki Kajiura; Yukitsugu Ono; Koichi Ishii; Akiko Tanaka; Junko Shinozaki; Yasuko Jojima
- Web page: https://patentgrep.com/patent/US7608695B2

## CPC

- C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)}
- C07K 14/755: Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)}
- G01N 33/86: involving blood coagulating time {or factors, or their receptors}
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2319/00: Fusion polypeptide
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- G01N 2333/755: Factors VIII, e.g. factor VIII C [AHF], factor VIII Ag [VWF]
- 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)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A method for conveniently detecting binding between the von Willebrand factor and glycoprotein Ib and a means to be used therein. The von Willebrand factor fixed in a reactor immobilized in a reaction vessel in the presence of bottrocetin is bound to a chimeric protein constructed by fusing the carboxyl terminal of a partial protein containing the von Willebrand factor-binding site of glycoprotein Ib with the amino terminal of the Fc region of an immunoglobulin molecule. Then the Fc region of the above immunoglobulin molecule is detected to thereby detect the binding between the von Willebrand factor and the glycoprotein Ib or inhibition of this binding.

## Claims

### Claim 1

A chimeric protein, which consists of an Fc region of immunoglobulin molecule fused at its amino terminus to a partial protein of glycoprotein Ibα chain at its carboxyl terminus,
wherein said partial protein consists of amino acid residues 1-319 of glycoprotein Ibα chain, and
wherein the immunoglobulin is derived from human.

### Claim 2

A kit for measuring glycocalicin based on inhibition of a reaction of von Willebrand factor and glycoprotein Ib, comprising von Willebrand factor and a chimeric protein that consists of an Fc region of immunoglobulin molecule fused at its amino terminus to a partial protein of glycoprotein Ibα chain at its carboxyl terminus,
wherein said partial protein consists of amino acid residues 1-319 of glycoprotein Ibα chain.

### Claim 3

A chimeric protein, which consists of an Fc region of immunoglobulin molecule fused at its amino terminus to a partial protein of glycoprotein Ibα chain at its carboxyl terminus,
wherein said partial protein consists of amino acid residues 1-293 of glycoprotein Ibα chain, and
wherein the immunoglobulin is derived from human.

### Claim 4

A kit for measuring glycocalicin based on inhibition of a reaction of von Willebrand factor and glycoprotein Ib, comprising von Willebrand factor and a chimeric protein that consists of an Fc region of immunoglobulin molecule fused at its amino terminus to a partial protein of glycoprotein Ibα chain at its carboxyl terminus,
wherein said partial protein consists of amino acid residues 1-293 of glycoprotein Ibα chain.

### Claim 5

A chimeric protein, which consists of an Fc region of immunoglobulin molecule fused at its amino terminus to a partial protein of glycoprotein Ibα chain at its carboxyl terminus,
wherein said partial protein consists of amino acid residues 251-285 of glycoprotein Ibα chain, and
wherein the immunoglobulin is derived from human.

### Claim 6

A kit for measuring glycocalicin based on inhibition of a reaction of von Willebrand factor and glycoprotein Ib, comprising von Willebrand factor and a chimeric protein that consists of an Fc region of immunoglobulin molecule fused at its amino terminus to a partial protein of glycoprotein Ibα chain at its carboxyl terminus,
wherein said partial protein consists of amino acid residues 251-285 of glycoprotein Ibα chain.