Highly active silica magnetic nanoparticles for purifying biomaterial and preparation method thereof
Inventors
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Assignees
Member
BioneerBioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Bioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Publication Number
US-10465184-B2
Publication Date
2019-11-05
Expiration Date
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.
Core Innovation
The invention relates to a method for preparing silica-coated magnetic nanoparticles by ultrasonically dispersing magnetic nanoparticles with an acid catalyst or base catalyst, a hydrophobic organic solvent, water, and an alcohol, and then adding silane to the dispersed solution to prepare silica magnetic nanoparticles. The silane is selected from tetramethoxysilane or tetraethoxysilane, and is added in a specified amount relative to the total volume of water, alcohol, catalyst, and hydrophobic organic solvent. The magnetic nanoparticles are also added in a specified amount relative to the total weight of the water, alcohol, catalyst, and hydrophobic organic solvent, and the magnetic nanoparticles and the silane are added at a defined weight ratio.
The resulting silica-coated magnetic nanoparticles are defined by a silica content of 0.5–2.5 wt%, a particle size of 300–600 nm, and a coating thickness of 1–15 nm, with a silica surface having hydroxyl groups. In the dispersing step, the hydrophobic organic solvent is selected from pentane, cyclopentane, hexane, cyclohexane, toluene, benzene, xylene, diethyl ether, dioxane, chloroform, and dichloromethane, and the alcohol is selected from methyl alcohol, ethyl alcohol, propyl alcohol, and butyl alcohol. The acid catalyst or base catalyst is selected from ammonia water, ammonium fluoride, hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, fluoric acid, oxalic acid, and acetic acid, and the magnetic nanoparticles include one or more selected from iron, cobalt, nickel, iron oxides, cobalt oxides, nickel oxides, and mixtures thereof.
The disclosed silica-coated magnetic nanoparticles are used as nucleic-acid-binding carriers for nucleic-acid isolation and purification with chaotropic-reagent-based binding and magnetic-force collection and subsequent elution. The document further describes nucleic-acid purification and nucleic-acid diagnosis kit/composition concepts associated with such nucleic-acid purification. The examples compare nucleic-acid extraction performance of these particles with conventional silica magnetic beads, including higher DNA binding capacity and electrophoresis results across different DNA sizes.
Claims Coverage
The document provides one independent claim directed to a preparation method for silica-coated magnetic nanoparticles, and it defines the inventive structure through specified dispersing components and silane addition conditions, resulting in defined particle and coating characteristics. Additional inventive refinements are presented in dependent claims that constrain formulation variables and nanoparticle selection, and that specify particular reagent sets as embodiments.
Ultrasonic dispersion with defined solvent/catalyst composition and silane addition to form silica magnetic nanoparticles
Ultrasonically dispersing a solution consisting of magnetic nanoparticles, an acid catalyst or base catalyst, a hydrophobic organic solvent, water, and an alcohol; adding silane to prepare silica magnetic nanoparticles; wherein the silane is added in an amount of 0.01–0.5 parts by volume based on 100 parts by volume of total water, alcohol, catalyst, and hydrophobic organic solvent; wherein the magnetic nanoparticles are added in 0.01–0.5 parts by weight based on 100 parts by weight of total water, alcohol, catalyst, and hydrophobic organic solvent; and wherein the magnetic nanoparticles and the silane are added at a weight ratio of 1:0.1–1:3.0.
Defined silica-coated magnetic nanoparticle characteristics with hydroxyl-functionalized silica surface
Silica-coated 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, wherein a surface of the silica has a hydroxyl group thereon.
Specified selections of hydrophobic solvent, alcohol, acid/base catalyst, silane, and magnetic nanoparticle species
Hydrophobic organic solvent selected from pentane, cyclopentane, hexane, cyclohexane, toluene, benzene, xylene, diethyl ether, dioxane, chloroform, and dichloromethane; alcohol selected from methyl alcohol, ethyl alcohol, propyl alcohol, and butyl alcohol; silane as tetramethoxysilane or tetraethoxysilane; acid catalyst or base catalyst selected from ammonia water, ammonium fluoride, hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, fluoric acid, oxalic acid, and acetic acid; and magnetic nanoparticles selected from iron, cobalt, nickel, iron oxides, cobalt oxides, nickel oxides, and mixtures thereof.
Across the independent claim, the inventive subject matter is a method that uses ultrasonic dispersion with a defined hydrophobic solvent/water/alcohol and acid/base catalyst system, followed by silane addition at defined amounts and ratios, to obtain silica-coated magnetic nanoparticles with defined silica content, particle size, coating thickness, and hydroxyl-functionalized silica surface. Dependent claims further constrain the water fraction in the dispersing solution and relate silica coating thickness to the silane addition amount, and narrow the nanoparticle selection to specified iron oxide/ferrite species; further dependent claims specify particular embodiment reagent sets.
Stated Advantages
Higher DNA binding capacity than conventional silica magnetic beads.
Improved nucleic-acid extraction performance with electrophoresis results across different DNA sizes.
Documented Applications
Nucleic-acid isolation and purification using chaotropic-reagent-based binding, magnetic-force collection, and elution, employing the silica-coated magnetic nanoparticles as nucleic-acid-binding carriers.
Nucleic-acid diagnosis using nucleic-acid purification and nucleic-acid diagnosis kit/composition concepts based on the nucleic-acid purification approach.
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