HIGHLY ACTIVE SILICA MAGNETIC NANOPARTICLES FOR PURIFYING BIOMATERIAL AND PREPARATION METHOD THEREOF

• US 20160348094A1
• Filed: 09/15/2015
• Published: 12/01/2016
• Est. Priority Date: 05/28/2015
• Status: Active Grant

• Current Assignee: Bioneer Corporation
• Original Assignee: Bioneer Corporation
• Inventors: Park, Jong Gwang, Park, Han Oh, Kim, Jae Ha
• Granted Patent: US 10,465,184 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: B22F 1/056 Submicron particles having a size above 100 nm up to 300 nm
• CPC Class Codes: B22F 1/105 containing inorganic lubricating or binding agents, e.g. metal salts
• CPC Class Codes: B22F 1/147 Making a dispersion
• CPC Class Codes: B22F 1/148 Agglomerating
• CPC Class Codes: B22F 1/16 Metallic particles coated with a non-metal coated with lubricating or binding agents or with organic material B22F1/10
• CPC Class Codes: B22F 2999/00 Aspects linked to processes or compositions used in powder metallurgy
• CPC Class Codes: C01B 33/18 Preparation of finely divided silica neither in sol nor in gel form; After-treatment thereof preparation of aerogels by dehydrating gels C01B33/158; treatment to enhance the pigmenting or filling properties C09C
• CPC Class Codes: C01P 2004/64 Nanometer sized, i.e. from 1-100 nanometer
• CPC Class Codes: C01P 2006/42 Magnetic properties
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: H01F 1/0054 Coated nanoparticles, e.g. nanoparticles coated with organic surfactant
• CPC Class Codes: H01F 1/06 in the form of particles, e.g. powder H01F1/047 takes precedence ; record carriers G11B5/70605

Abstract

The present invention relates to a method for preparing highly active silica magnetic nanoparticles, highly active silica magnetic nanoparticles prepared by the method, and a method of isolating nucleic acid using the highly active silica magnetic nanoparticles. The highly active silica magnetic nanoparticles prepared according to the present invention contain magnetic nanoparticles completely coated with silica, can be used as a reagent for isolating biomaterials, particularly, nucleic acids, and can isolate and purify nucleic acid in a high yield.

Claims

1. A method for preparing highly active silica magnetic nanoparticles comprising:

(a) ultrasonically dispersing a solution obtained by adding magnetic nanoparticles, an acid-base catalyst and a water-soluble hydrophobic solvent to a solvent mixture of distilled water and an alcohol; and

(b) adding silane to the dispersed solution to prepare silica magnetic nanoparticles.

2. The method of claim 1, wherein the alcohol is selected from the group consisting of methyl alcohol, ethyl alcohol, propyl alcohol, and butyl alcohol.

3. The method of claim 1, wherein the solvent mixture contains 30 vol % or less of distilled water.

4. The method of claim 1, wherein the silane is tetramethoxysilane or tetraethoxysilane.

5. The method of claim 1, wherein the acid-base catalyst is selected from the group consisting of ammonia water (NH₄OH), ammonium fluoride (NH₄F), hydrochloric acid (HCl), nitric acid (HNO₃), sulfuric acid (H₂SO₄), phosphoric acid (H₃PO₄), fluoric acid (HF), oxalic acid, and acetic acid.

6. The method of claim 1, wherein the silane is added in an amount of 0.01-0.5 parts by volume based on 100 parts by volume of the solvent.

7. The method of claim 6, wherein a thickness of silica coated on a surface of the silica magnetic nanoparticles is controlled according to an added amount of silane.

8. The method of claim 1, wherein the magnetic nanoparticles are one or more selected from the group consisting of iron, cobalt, nickel, iron oxides, cobalt oxides, nickel oxides, and mixtures thereof.

9. The method of claim 8, wherein the magnetic nanoparticles are one or more selected from the group consisting of iron oxide, ferrite, and mixtures thereof.

10. The method of claim 7, wherein the magnetic nanoparticles are added in an amount of 0.01-0.5 parts by weight based on 100 parts by weight of distilled water, alcohol, the acid-base catalyst and the water-soluble hydrophobic solvent.

11. The method of claim 1, wherein the water-soluble hydrophobic solvent is selected from the group consisting of pentane, cyclopentane, hexane, cyclohexane, toluene, benzene, xylene, diethyl ether, dioxane, chloroform, and dichloromethane

12. The method of claim 1, wherein the magnetic nanoparticles and the silane are added at a weight ratio of 1:

0.1-1;

3.0.

13. Highly active silica magnetic nanoparticles having a silica content of 0.5-2.5 wt %, a particle size of 300-600 nm, and a coating thickness of 1-15 nm.

14. Highly active silica magnetic nanoparticles of claim 13, which are prepared by the method comprising:

(a) ultrasonically dispersing a solution obtained by adding magnetic nanoparticles, an acid-base catalyst and a water-soluble hydrophobic solvent to a solvent mixture of distilled water and an alcohol; and

(b) adding silane to the dispersed solution to prepare silica magnetic nanoparticles.

15. A method for isolating and purifying nucleic acid, wherein the highly active silica magnetic nanoparticles of claim 13 are used as nucleic acid-binding carriers.

16. The method of claim 15, wherein nucleic acid is selected from the group consisting of plasmid DNA, genomic DNA, cDNA, PCR DNA (polymerase chain reaction DNA), RNA, siRNA, ribozymes, aptamers, and oligonucleotide.

17. A composition for purifying nucleic acid, which comprises the highly active silica magnetic nanoparticles of claim 13.

18. A composition for diagnosing by nucleic acid detection, which comprises the highly active silica magnetic nanoparticles of claim 13.

19. A kit for purifying nucleic acid, which comprises the highly active silica magnetic nanoparticles of claim 13.

20. A kit for diagnosing nucleic acid detection, which comprises the highly active silica magnetic nanoparticles of claim 13.

21. A method wherein the highly active silica magnetic nanoparticles of claim 13 are used in a process selected from the group consisting of protein purification, antibody purification, enzyme immunoassay, peptide purification, and endotoxin removal.