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Surface functionalization of gold or silver nanoparticle, and colorimetry detection method for molecule by using the same

A silver nanoparticle and surface functionalization technology, which is applied in biological testing, color/spectral characteristic measurement, material inspection products, etc., can solve the problems of difficult realization of gold nanoparticle labeling, achieve easy purification and quantitative analysis, and avoid detection equipment , the effect of chemical stability

Inactive Publication Date: 2006-07-19
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thilylated oligonucleotides are commercially available, but most other molecules that are thiolated are not commercially available, and therefore, labeling of other molecules with gold nanoparticles by this method is difficult.
The method of labeling DNA with other kinds of metal nanoparticles has not been reported

Method used

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  • Surface functionalization of gold or silver nanoparticle, and colorimetry detection method for molecule by using the same
  • Surface functionalization of gold or silver nanoparticle, and colorimetry detection method for molecule by using the same
  • Surface functionalization of gold or silver nanoparticle, and colorimetry detection method for molecule by using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1: Determination of DNA by gold nanoparticle-labeled oligonucleotide probes

[0015] Coupling of oligonucleotides to dextran: 0.16ml, 25mg / ml 4-nicotinic acid succinimide ester hydrazine acetone hydrazone (C6-succinimidyl 4-hydrazinonicotinate acetone hydrazone) and 25mg / ml N-succinyl Amine-3-(2-pyridyldithio)-propionate (SPDP, N-succinimidyl3-(2-pyridyldithio)propionate) in anhydrous DMSO solution was added to 4ml, 2mg / ml aminodextran (MW 70KDa ) in 1×PBS solution at room temperature for 2 hours. The resulting dextran conjugate was purified on a Sephadex G25 column using 0.1M NaAc (pH=4.5) as the eluent. The amount of dithio and hydrazone functional groups generated can be quantified with 1,4-dithiothreitol and 4-nitrobenzaldehyde, respectively. Excess aldehyde-modified oligonucleotides were added to a 0.1M NaAc (pH4.5) solution of dextran conjugates and reacted for 12 hours, and the unreacted oligonucleotides were separated and removed with a 30KDa molecular...

Embodiment 2

[0018] Example 2: Silver nanoparticle-labeled oligonucleotide probes measure DNA

[0019]Coupling of oligonucleotides to dextran: 0.16ml, 25mg / ml 4-nicotinic acid succinimide ester hydrazine acetone hydrazone (C6-succinimidyl 4-hydrazinonicotinate acetone hydrazone) and 25mg / ml N-succinyl Amine-3-(2-pyridyldithio)-propionate (SPDP, N-succinimidyl3-(2-pyridyldithio)propionate) in anhydrous DMSO solution was added to 4ml, 2mg / ml aminodextran (MW 70KDa ) in 1×PBS solution at room temperature for 2 hours. The resulting dextran conjugate was purified on a Sephadex G25 column using 0.1M NaAc (pH=4.5) as the eluent. The amount of dithio and hydrazone functional groups generated can be quantified with 1,4-dithiothreitol and 4-nitrobenzaldehyde, respectively. Excessive CHO-modified oligonucleotide probes were added to 0.1M NaAc (pH4.5) solution of dextran conjugates and reacted for 12h, then separated and removed unreacted oligonucleotide probes with a 30KDa molecular weight cut-off ...

Embodiment 3

[0022] Example 3: Gold Nanoparticles Label Biotin Molecules

[0023] Conjugation of biotin to dextran: 5 mg N-succinimide-3-(2-pyridyldithio)-propionate (SPDP) dissolved in 100 μl, 50 mg / ml biotinaminocaproic acid-N- Hydroxysuccinimide ester (Biotinamidohexanic acid N-hydroxysuccinimide ester) anhydrous DMSO solution, this solution was added dropwise to 2.5ml, 2mg / ml aminodextran (MW 70KDa) NaHCO 3 (50mM) solution at room temperature for 2 hours. The resulting dextran conjugates are purified by dialysis in aqueous solution for further use. The amount of biotin on the dextran conjugate can be quantified with HABA (4'-hydroxyazobenzene-2-carboxylic acid).

[0024] Preparation of gold nanoparticle-labeled biotin-dextran conjugate probe: add different amounts of biotin-dextran conjugate to a certain amount of gold nanoparticle solution, and record the solution after filtering through a 0.20 μm membrane According to the UV absorption of gold nanoparticles at 520nm, the desired a...

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Abstract

The invention relates to a method for labeling and colorimetric detecting the biology molecule by surface fictionalized gold or silver nanometer particle. The probe molecule for identifying the biology molecule and the nail molecule for connecting the gold or silver nanometer particle first through the gluglucosan whose double function agent is covalence reformatted macromolecule to form the deformation coated the gold or silver nanometer particle, it uses the self-component on the surface of the gold or silver nanometer particle by the nail group of the deformation to form gluglucosan molecule coated gold or silver nanometer particle after purifying and quantifying the deformation.

Description

technical field [0001] The invention relates to a method for colorimetrically detecting biomolecules using surface-functionalized gold or silver nanoparticles as markers, and belongs to the technical field of nano-labeling and detection of biomolecules. Background technique [0002] Label analysis is one of the most important methods for the detection of biomolecules (nucleic acid, protein, etc.). Due to the weak nature of many biomolecules that can be analyzed by themselves, in order to obtain high-sensitivity detection, it is often analyzed with the help of external markers to obtain measurable signals. Commonly used labels can be divided into luminescent labels, enzyme labels, electrochemical labels and so on. Among them, the luminescent labeling method is the most important method, such as various fluorescent labels and chemiluminescent labels. The sensitivity of luminescent label detection depends largely on the luminescent intensity of the luminescent label. [0003...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/25
Inventor 陈扬
Owner SOUTHEAST UNIV
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