Human resistin receptor and use thereof

Patent metadata

Field	Value
Publication number	US-9933424-B2
Application number	US-201214237518-A
Country	US
Kind code	B2
Filing date	Aug 7, 2012
Priority date	Aug 8, 2011
Publication date	Apr 3, 2018
Grant date	Apr 3, 2018

Abstract

The present invention concerns a human resistin receptor. More particularly, the present invention provides a method for screening a receptor of human resistin protein, a method for preventing or treating an inflammatory disease and arteriosclerosis using an expression- or activity-regulator for a human resistin receptor, and a pharmaceutical composition including an expression- or activity-regulator for the human resistin receptor. The method for screening a human resistin protein receptor according to the present invention enables separation of a receptor which directly binds to resistin from human monocyte, reveals a mechanism of signal transduction of the resistin receptor, and therefore, is expected to contribute to regulation of an inflammatory effect of monocyte, molecular detection of causes for vascular inflammation and arteriosclerosis, and developments of prevention and a treating agent for an inflammatory disease and arteriosclerosis.

First claim

The invention claimed is:

1. A method for screening a human resistin protein receptor comprising:
a) a step of preparing a recombinant vector by cloning mFc-human resistin recombinant DNA to an expression vector, wherein the mFc-human resistin recombinant DNA comprises a nucleic acid encoding an mFc human resistin fusion protein;
b) a step of expressing the mFc human resistin fusion protein by transfecting the recombinant vector to a cell strain;
c) a step of forming a complex of the expressed mFc human resistin fusion protein and a human resistin receptor by cultivating the expressed mFc human resistin fusion protein with human acute monocytic leukemia (THP-1) cells;
d) a step of immuno-precipitating the complex and separating the human resistin receptor from the precipitate; and
e) a step of confirming the separated human resistin receptor.

2. The method according to claim 1 , wherein the expression vector in the step a) is pcDNA3.1.

3. The method according to claim 1 , wherein the cell strain in the step b) is a HEK293F cell.

4. The method according to claim 1 , further comprising a step of purifying the expressed mFc human resistin fusion protein.

5. The method according to claim 1 , wherein the expressed mFc human resistin fusion protein is cultivated together with an anti-mFc-FITC secondary antibody in the step c).

6. The method according to claim 1 , wherein beads specific to mFc are used in the immuno-precipitation in the step d).

7. The method according to claim 1 , wherein the human resistin receptor in the step d) is a protein having a size of 55 kDa.

8. A method for screening a human resistin protein receptor comprising:
a) a step of preparing a recombinant vector by cloning mFc-human resistin recombinant DNA to an expression vector, wherein the mFc-human resistin recombinant DNA comprises a nucleic acid encoding an mFc human resistin fusion protein;
b) a step of expressing mFc human resistin fusion protein by transfecting the recombinant vector to an HEK293F cell;
c) a step of purifying the expressed mFc human resistin fusion protein;
d) a step of forming a complex of the expressed mFc human resistin fusion protein and a human resistin receptor by cultivating the purified mFc human resistin fusion protein together with a THP-1 cell;
e) a step of immuno-precipitating the complex to obtain a precipitate using beads specific to mFc;
f) a step of separating the human resistin receptor corresponding to a size of 55 kDa from the precipitate; and
g) a step of confirming the separated human resistin receptor by mass spectrometry.

Assignees

• Park Young Bae
• Kim Hyo Soo
• Kwon Yoo Wook
• Lee Sahmin
• Lee Hyun Chae
• Cho Young Jin
• Lee Sang Eun
• Chung Jun Ho
• Seoul Nat Univ Hospital

Inventors

• Park Young Bae
• Kim Hyo Soo
• Kwon Yoo Wook
• Lee Sahmin
• Lee Hyun Chae
• Cho Young Jin
• Lee Sang Eun
• Chung Jun Ho

Classifications

• G01N33/6863

  Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors · CPC title

• G01N2333/715

  for cytokines; for lymphokines; for interferons · CPC title

• G01N33/564 Primary

  for pre-existing immune complex or autoimmune disease {, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9} · CPC title

• C12N15/85

  for animal cells · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title