Nanoparticles in the shape of nanosnowman with a head part and a body part, a preparation method thereof and a detection method using the same

Abstract

The present invention relates to nanoparticles in the shape of nanosnowman with a head part and a body part, a preparation method thereof, and a detection method using the same. More particularly, the present invention relates to nanoparticles in the shape of nanosnowman with head and body parts, which can offer platforms for DNA-based assembly of various aligned and unconventional nanostructures and is highly applicable to the detection of DNA and an analyte associated with the onset and progression of a particular disease, a preparation method thereof, and a detection method using the same.

1 . Nanoparticles that are composed of a gold or silver nanoparticle head part and a gold or silver nanoparticle body part, wherein a plurality of oligonucleotides are bound to the surface of the head part, and a lower portion of the head part is located on a concave region in the upper portion of the body part. 
     
     
         2 . The nanoparticles according to  claim 1 , wherein the head part has a diameter of 2 to 200 nm. 
     
     
         3 . The nanoparticles according to  claim 1 , wherein the body part has a diameter of 2 to 900 nm. 
     
     
         4 . The nanoparticles according to  claim 1 , wherein the head part and the body part are asymmetric in size. 
     
     
         5 . The nanoparticles according to  claim 1 , wherein the longest axis of the nanoparticle is 4 nm to 900 nm. 
     
     
         6 . The nanoparticles according to  claim 1 , wherein the head part has a shape of sphere, nanorod or nanocube. 
     
     
         7 . The nanoparticles according to  claim 1 , wherein the body part has a shape of sphere, nanorod or nanocube. 
     
     
         8 . The nanoparticles according to  claim 1 , wherein the body part has a hollow structure. 
     
     
         9 . The nanoparticles according to  claim 1 , wherein a part of the oligonucleotides bound to the surface of the head part are exposed outside and the rest thereof are buried in a concave region in the upper portion of the body part, and consequently, the nanoparticle have asymmetrically modified oligonucleotides. 
     
     
         10 . The nanoparticles according to  claim 1 , wherein the oligonucleotides are bound to the surface of the gold or silver head part by any one functional group selected from the group consisting of a thiol group, an amino group, and an alcohol group. 
     
     
         11 . The nanoparticles according to  claim 10 , wherein the oligonucleotide includes a spacer sequence between the functional group and the oligonucleotide. 
     
     
         12 . The nanoparticles according to  claim 11 , wherein the spacer sequence is represented by -PEG x -Y y -(CH 2 ) z -, x is an integer of 0 to 30, y is an integer of 0 to 30, z is an integer of 3 to 6, and Y is adenine, thymine, guanine or cytosine, respectively. 
     
     
         13 . The nanoparticles according to  claim 12 , wherein the spacer sequence is any one selected from the group consisting of PEG 18 -A 10 -(CH 2 ) 3 , PEG 18 -A 10 -(CH 2 ) 6 , PEG 18 -A 30 -(CH 2 ) 3 , PEG 18 -A 30 -(CH 2 ) 6 , PEG 18 -T 10 -(CH 2 ) 3 , PEG 18 -T 10 -(CH 2 ) 6 , PEG 18 -T 30 -(CH 2 ) 3 , PEG 18 -T 30 -(CH 2 ) 6 , A 10 -(CH 2 ) 3 , A 10 -(CH 2 ) 6 , A 30 -(CH 2 ) 3 , A 30 -(CH 2 ) 6 , T 10 -(CH 2 ) 3 , T 10 -(CH 2 ) 6 , T 30 -(CH 2 ) 3 , T 30 -(CH 2 ) 6 , PEG-A 10 , PEG-A 10 , PEG-A 30 , PEG-A 30 , PEG-T 10 , PEG-T 10 , PEG-T 30 , and PEG-T 30 . 
     
     
         14 . The nanoparticles according to  claim 1 , wherein a Raman active molecule binds to the oligonucleotide. 
     
     
         15 . The nanoparticles according to  claim 14 , wherein the Raman active molecule is selected from the group consisting of FAM, Dabcyl, TRIT (tetramethyl rhodamine isothiol), NBD (7-nitrobenz-2-1,3-diazole), Texas Red dye, phthalic acid, terephthalic acid, isophthalic acid, cresyl fast violet, cresyl blue violet, brilliant cresyl blue, para-aminobenzoic acid, erythrosine, biotin, digoxigenin, 5-carboxy-4′,5′-dichloro-2′,7′-dimethoxy, fluorescein, 5-carboxy-2′,4′,5′,7′-tetrachlorofluorescein, 5-carboxyfluorescein, 5-carboxyrhodamine, 6-carboxyrhodamine, 6-carboxytetramethyl aminophthalocyanine, azomethine, cyanine, xanthine, succinylfluorescein, aminoacridine, quantum dots, carbon nanotubes, carbon allotropes, cyanide, thiol, chlorine, bromine, methyl, phosphorus, sulfur, cyanine dyes (Cy3, Cy3.5, Cy5), and rhodamine. 
     
     
         16 . A method for preparing the nanoparticles of  claim 1 , comprising the following steps of:
 1) modifying a gold or silver nanoparticle with oligonucleotides (step 1); and   2) reacting the oligonucleotide-modified gold or silver nanoparticle with a gold or silver precursor in the presence of NaCl, a reducing agent, and a stabilizer (step 2).   
     
     
         17 . The method according to  claim 16 , wherein the gold or silver precursor is AgNO 3 , AgClO 4  or HAuCl 4 . 
     
     
         18 . The method according to  claim 16 , wherein the NaCl concentration is  1  nM to  0 . 1  M. 
     
     
         19 . The method according to  claim 16 , wherein step 2) is performed under the condition of pH 2 to 7. 
     
     
         20 . The method according to  claim 16 , wherein the reducing agent is hydroquinone, sodium borohydride (NaBH 4 ), sodium ascorbate, hydroxyl amine or a combination thereof. 
     
     
         21 . The method according to  claim 16 , wherein the stabilizer is a derivative including pyrrolidine, imidazolidine, pyrazolidine, piperidine, piperazine, sorbitol, ethylene glycol or a carbonyl group; saccharose including glucose or fructose; DNA; PNA; or RNA. 
     
     
         22 . The method according to  claim 16 , wherein the stabilizer is polyvinylpyrrolidone (PVP). 
     
     
         23 . A method for detecting an analyte, comprising the steps of:
 functionalizing the surface of the nanoparticle of  claim 1  with a biomolecule capable of recognizing the analyte to be analyzed;   exposing the nanoparticle to a sample containing one or more analytes; and   detecting and identifying one or more analytes by laser excitation and Raman spectroscopy.   
     
     
         24 . The method according to  claim 23 , wherein the Raman spectroscopy is Surface Enhanced Raman Scattering (SERS), Surface enhanced resonance Raman spectroscopy (SERRS), or hyper-Raman and/or Coherent Anti-Stokes Raman Spectroscopy (CARS). 
     
     
         25 . A kit for detecting an analyte, comprising the nanoparticles of  claim 1 . 
     
     
         26 . A molecular diagnostic chip or a composition for diagnostic imaging, comprising the nanoparticles  claim 1 . 
     
     
         27 . Nanoparticles, comprising the nanoparticles of  claim 1 ; and further any one selected from the group consisting of a CT contrast agent, an MRI contrast agent, an optical imaging contrast agent, and an ultrasound contrast agent inside or outside thereof. 
     
     
         28 . Nanoparticles, comprising the nanoparticles of  claim 1 ; and further any one selected from the group consisting of genes, antibodies, and drugs.