# US 9,751,910 B2 · Method for preparing metal nanostructure based on biomolecules

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- Number: US 9,751,910 B2
- Document ID: US9751910B2
- Status: Granted
- Kind: B2
- Application number: 14/898,741
- Filed: 2014-07-03
- Published: 2016-05-19
- Granted: 2017-09-05
- Earliest claimed: 2013-08-13
- Assignees: Korea University Research and Business Foundation
- Applicants: Korea University Research and Business Foundation
- Inventors: Sang Jun Sim; Xingyi Ma; Jong-Uk Lee
- Web page: https://patentgrep.com/patent/US9751910B2

## CPC

- C07H 23/00: Compounds containing boron, silicon or a metal, e.g. chelates or vitamin B12 (esters with inorganic acids C07H11/00)
- A61K 9/5115: {Inorganic compounds}
- A61K 9/5192: {Processes}
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- B22F 1/054: Nanosized particles
- B22F 1/102: Metallic powder coated with organic material
- B22F 9/24: starting from liquid metal compounds, e.g. solutions
- B82B 3/0033: {Manufacture or treatment of substrate-free structures, i.e. not connected to any support}
- G01N 21/554: {detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance}
- G01N 27/44747: {Composition of gel or of carrier mixture}
- B82Y 40/00: Manufacture or treatment of nanostructures
- C22C 2202/02: Magnetic
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- 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)
- B22F: WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER (making alloys by powder metallurgy C22C); APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B82B: NANOSTRUCTURES FORMED BY MANIPULATION OF INDIVIDUAL ATOMS, MOLECULES, OR LIMITED COLLECTIONS OF ATOMS OR MOLECULES AS DISCRETE UNITS; MANUFACTURE OR TREATMENT THEREOF
- 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)
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- C22C: ALLOYS (treatment of alloys C21D, C22F)

## Abstract

The present invention relates to a method for preparing metal nanostructures using DNA, and more particularly, to a method for preparing metal nanostructures, in which a self-assembling DNA is used as a frame, and thus the orientation, shape and size of the nanostructures are easily controlled compared to conventional bottom-up methods. Metal nanostructures prepared by the method show excellent localized surface plasmon resonance properties, and thus can be used as fluorescent substances in drug delivery, biomedical imaging, and supersensitive biosensors.

## Claims

### Claim 1

A method for preparing a biomolecule-based metal nanostructure, the method comprising the steps of:
(a) forming a self-assembled monomolecular layer of a biomolecule on the surface of a metal nanoseed, thereby forming a metal nanoseed-biomolecule complex; and
(b) growing a metal ion on the surface of the complex while reducing the metal ion with a reducing agent,
wherein the metal nanoseed-biomolecule complex is prepared by the steps of:
(i) binding a desthiobiotin-modified DNA to a streptavidin-modified magnetic particle to form a magnetic particle-DNA complex;
(ii) binding the metal nanoseed to the DNA of the complex to form a magnetic particle-DNA-metal nanoseed complex; and
(iii) adding a biotin solution to the magnetic particle-DNA-metal nanoseed complex to separate the bond between the magnetic particle and the DNA and remove the magnetic particle from the complex, thereby obtaining a metal nanoseed-DNA complex,
wherein steps (i) to (iii) are repeatedly performed.

### Claim 2 (depends on 1)

The method of claim 1, wherein step (i) further comprises, after forming the magnetic particle-DNA complex, a step of adding the magnetic particle-DNA complex to a mixture solution of EDC and NHS to activate the DNA.

### Claim 3

A method for preparing a biomolecule-based metal nanostructure, the method comprising the steps of:
(a) forming a self-assembled monomolecular layer of a biomolecule on the surface of a metal nanoseed, thereby forming a metal nanoseed-biomolecule complex; and
(b) growing a metal ion on the surface of the complex while reducing the metal ion with a reducing agent,
wherein the metal nanoseed-biomolecule complex is prepared by the steps of:
(i) treating the metal nanoseed surface with a dATP having a terminal disulfide group and an oligo(ethylene glycol) (OEG-OH) having a hydroxyl group to form a self-assembled monomolecular layer, thereby modifying the metal nanoseed surface;
(ii) adding a thiol-terminated DNA to the metal nanoseed, and separating and removing the dATP from the metal nanoseed by a ligand exchange reaction, thereby preparing a metal nanoseed-DNA complex; and
(iii) recovering the metal nanoseed-DNA complex by electrophoresis.

### Claim 4

A method for preparing a biomolecule-based metal nanostructure, the method comprising the steps of:
(a) forming a self-assembled monomolecular layer of a biomolecule on the surface of a metal nanoseed, thereby forming a metal nanoseed-biomolecule complex; and
(b) growing a metal ion on the surface of the complex while reducing the metal ion with a reducing agent,
wherein the metal nanoseed-biomolecule complex is prepared by the steps of:
(i) inserting a DNA intercalating molecule into a plasmid DNA, followed by desalting to obtain a plasmid DNA solution;
(ii) adding a metal nanoseed aqueous solution to the plasmid DNA solution to form a metal nanoseed-plasmid DNA complex; and
(iii) treating the metal nanoseed- plasmid DNA complex with a buffer (pH 10-14).

### Claim 5 (depends on 4)

The method of claim 4, wherein the DNA intercalating molecule is 1-pyrenebutyric acid (PBA) or S-(2-[[4-(2-phenanthryl)butyl]amino]ethyl)hydrogen sulfurothioate).

### Claim 6 (depends on 1, 3)

The method of any one of claims 1, 3, and 4, wherein the source of the metal ion is one selected from the group consisting of hydrogen tetrachloroaurate(III) (HAuCl4), sodium tetrachloroaurate(III) (NaAuCl4), gold(III) chloride (AuCl3), and potassium gold(III) chloride (KAuCl4).

### Claim 7 (depends on 1, 3)

The method of any one of claims 1, 3, and 4, wherein the reducing agent is one selected from the group consisting of hydroxylamine (NH2OH), sodium diphenylamine sulfonate, ascorbic acid, and poly(allylamine) hydrochloride.

### Claim 8 (depends on 1, 3)

The method of any one of claims 1, 3, and 4, wherein the method further comprises, after step (b), a step of adding mercaptopropionic acid to the complex to terminate the growth.

### Claim 9 (depends on 1, 3)

The method of any one of claims 1, 3, and 4, wherein the metal nanoseed is any one selected from the group consisting of nanospheres, nanorods, nanoprisms and nanosheets, and has a size of 1-100 nm.

### Claim 10 (depends on 1, 3)

The method of any one of claims 1, 3, and 4, wherein the metal nanoseed is made of any one selected from the group consisting of gold (Au), silver (Ag), platinum (Pt), palladium (Pd), copper (Cu), silicon (Si), germanium (Ge), aluminum (Al), and metal oxides.

### Claim 11 (depends on 1, 3)

The method of any one of claims 1, 3, and 4, wherein the biomolecule is any one selected from the group consisting of single-stranded DNAs, double-stranded DNAs, DNA oligomers, RNA oligomers, plasmid DNAs, polypeptides, and proteins, in which the single-stranded DNAs, the double-stranded DNAs, the DNA oligomers and the RNA oligomers are 5-5,000 bp in length, and the plasmid DNAs are 800-10,000 bp in length.