Polypeptide selectively binding to immunoglobulin G of mouse or rabbit and use thereof

Patent metadata

Field	Value
Publication number	US-10995153-B2
Application number	US-201616074530-A
Country	US
Kind code	B2
Filing date	Sep 20, 2016
Priority date	Feb 1, 2016
Publication date	May 4, 2021
Grant date	May 4, 2021

Abstract

The present invention relates to a novel polypeptide that binds selectively to mouse or rabbit immunoglobulin G. More specifically, the present invention relates to a polypeptide that binds to mouse or rabbit immunoglobulin G, a polynucleotide encoding the polypeptide, an expression vector comprising the polynucleotide, a transformant introduced with the expression vector, a method of producing the polypeptide using the transformant, and a composition for immunoassay comprising the polypeptide. The novel peptide according to the present invention binds specifically to mouse or rabbit immunoglobulin G, can replace conventional expensive secondary immunoglobulin G, and can be used in various biological immunoassays. In addition, a conjugate of the polypeptide of the present invention and immunoglobulin G is useful for fabrication of various immunosensors/immunochips and for drug screening.

First claim

The invention claimed is:

1. A repebody comprising any one of the amino acid sequences represented by SEQ ID NOS: 4 to 6 and 13 to 15, the repebody binding selectively to mouse immunoglobulin G.

2. A polynucleotide encoding the repebody of claim 1 .

3. A recombinant vector comprising the polynucleotide of claim 2 .

4. A recombinant microorganism into which the polynucleotide of claim 2 or a recombinant vector comprising said polynucleotide is introduced.

5. A method for producing the repebody of claim 1 , wherein the method comprises:
(i) introducing into a host microorganism (a) a polynucleotide encoding a repebody comprising any one of the amino acid sequences represented by SEQ ID NOS: 4 to 6 and 13 to 15, the repebody binding selectively to mouse immunoglobulin G, or (b) a recombinant vector comprising said polynucleotide, to thereby produce a recombinant microorganism;
(ii) expressing the repebody by culturing the recombinant microorganism; and
(iii) recovering the expressed repebody.

6. A method for purifying a mouse immunoglobulin G antibody, wherein the method comprises the steps of:
(i) injecting a mixture comprising a mouse immunoglobulin G antibody into a column into which the repebody of claim 1 is adsorbed; and
(ii) eluting the antibody attached to the column of step (i).

7. A method for immobilizing mouse immunoglobulin G, wherein the method comprises the steps of:
(i) treating a surface of a solid substrate by attaching the repebody of claim 1 onto the solid substrate; and
(ii) binding mouse immunoglobulin G to the surface-treated solid substrate.

8. An immunosensor having a solid substrate surface-treated with the repebody of claim 1 , such that the repebody is attached to the solid substrate, and wherein mouse immunoglobulin G is immobilized to the solid substrate by binding to the repebody.

9. A method of detecting a substance having binding affinity for mouse immunoglobulin G, the method comprising the steps of:
(a) treating the immunosensor of claim 8 with a sample containing the substance having binding affinity for mouse immunoglobulin G; and
(b) determining whether the substance would bind to the immunosensor.

10. A composition for ELISA of mouse immunoglobulin G, the composition comprising the repebody of claim 1 .

11. A composition for Western blotting of mouse immunoglobulin G, the composition comprising the repebody of claim 1 .

12. A composition for immunohistochemical staining of mouse immunoglobulin G, the composition comprising the repebody of claim 1 , which has a fluorescent substance conjugated thereto.

13. The composition of claim 12 , wherein the fluorescent substance is a fluorescent dye, a tetracystein motif, a fluorescent protein, a fluorescent nanoparticle, or a quantum dot.

Assignees

• Korea Advanced Inst Sci & Tech

Inventors

• Kim Hak-Sung
• Jeong Sukyo
• Heu Woosung
• Kim Jong-Won
• Lee Joong-Jae

Classifications

• C07K14/195 Primary

  from bacteria · CPC title

• G01N33/531

  Production of immunochemical test materials · CPC title

• G01N33/54306

  Solid-phase reaction mechanisms · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C07K14/705

  Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} · CPC title