Virus-microbead complex and use thereof

Patent metadata

Field	Value
Publication number	US-10107805-B2
Application number	US-201314758126-A
Country	US
Kind code	B2
Filing date	Sep 2, 2013
Priority date	Dec 28, 2012
Publication date	Oct 23, 2018
Grant date	Oct 23, 2018

Abstract

The present invention relates to a virus-microbead complex including a microbead and a virus layer, in which linear viruses are bound individually to the surface of the microbead, and an immunoassay kit including the same.
The virus-microbead complex of the present invention is characterized in that the linear viruses are bound to the surface of the microbead so that the orientations of the linear viruses are regulated using the interaction of streptavidin-biotin introduced thereon, thereby providing a significantly increased volume to surface area ratio, increasing the number of antibodies or ligands capable of binding thereto, and as a result, mediating the binding of antibodies or ligands to a unit bead with high density, which eventually leads to an increased sensitivity in immunoassays, and an application into a suspension array. Additionally, it was confirmed that cardiac troponin I (cTnI) in serum can be detected up to 20 pg/mL by introducing a self-assembled monolayer (SAM) containing PEG to remove a non-specific adsorption.

First claim

What is claimed is:

1. A virus-microbead complex comprising:
(i) a microbead made of gold or coated with a thin gold film,
(ii) a self-assembled monolayer (SAM) on the surface of the microbead, which is composed of carboxyl-terminated hexa(ethylene glycol)undecane thiol bound to streptavidin or biotin, preventing non-specific binding on the surface of the microbead,
(iii) a virus layer having one end of each linear virus bound to the SAM on the surface of the microbead,
wherein the individual linear virus in the virus layer are independently bound to the SAM, and
wherein the binding between the SAM on the surface of the microbead and the linear viruses is mediated by the binding between streptavidin (SAV) and biotin to separate one linear virus to another linear virus in the virus layer.

2. The complex of claim 1 , wherein at least two capturers capable of capturing an analyte are bound to the exposed long axis surface of the virus in the virus layer.

3. The complex of claim 2 , wherein the capturer is selected from the group consisting of an antibody, an antigen, a ligand, and a receptor.

4. The complex of claim 1 , wherein the linear virus is filamentous bacteriophage fd phage or M13 phage.

5. The complex of claim 1 , wherein biotin or streptavidin is bound to a protein located at one end of the virus, or the virus is gene-manipulated to express a fusion protein of biotin or streptavidin and a virus protein.

6. The complex of claim 5 , wherein the protein located at one end of the virus is pIII, pVI, pVII, pIX, or a combination thereof.

7. The complex of claim 1 , wherein the microbead is magnetic.

8. An immunoassay kit comprising the virus-microbead complex of claim 1 .

9. The kit of claim 8 , comprising the complex; and further a capturer for capturing an analyte, which is bindable to the surface of the virus within the complex.

10. The kit of claim 8 , wherein the virus and the capturer for capturing an analyte are linked by a covalent bond.

11. The kit of claim 10 , wherein the covalent bond is achieved by Staudinger ligation.

12. The kit of claim 8 , wherein the kit is for suspension array.

13. The kit of claim 8 , wherein when the microbead is magnetic, the kit can separate an analyte by separating the complex bound to the analyte using a magnetic property.

14. The complex of claim 3 , wherein the antibody is modified with azide, and linked to the phosphine group bound to Ala or Lys residue located at N-terminal of the coat protein exposed to the surface of the linear virus.

Assignees

• Seoul Nat Univ R&Db Foundation

Inventors

• Chung Taek Dong
• Hwang Inseong
• Jeon Chang Su

Classifications

• G01N33/553

  Metal or metal coated · CPC title

• G01N33/554

  the carrier being a biological cell or cell fragment, e.g. bacteria, yeast cells · CPC title

• G01N33/54333 Primary

  Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction · CPC title

• G01N33/5432

  Liposomes or microcapsules · CPC title

• G01N2333/005

  from viruses · CPC title