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Electrochemical immunoassay method based on Au-PB-SiO2 composite nano-particles

An immunoassay method and composite nanoparticle technology, applied in the field of electrochemical immunity, can solve the problems affecting the accuracy of small cell lung cancer, the sensitivity of enzyme-linked immunoassay is not high, and it is difficult to detect trace neuron-specific enolase

Inactive Publication Date: 2013-12-11
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection of neuron-specific enolase (NSE) by enzyme-linked immunosorbent method at home and abroad has been in the clinical application stage, but the sensitivity of enzyme-linked immunosorbent assay is not high, and it is difficult to detect trace amounts of neuron-specific enolase (NSE) , affecting the accuracy of detection of small cell lung cancer (SCLC)

Method used

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  • Electrochemical immunoassay method based on Au-PB-SiO2 composite nano-particles
  • Electrochemical immunoassay method based on Au-PB-SiO2 composite nano-particles
  • Electrochemical immunoassay method based on Au-PB-SiO2 composite nano-particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1 prepares Au-PB-SiO 2 composite nanoparticles

[0062] Add 2 mL of tetraethyl orthosilicate (TEOS) into a mixed solution of 10 mL of ethanol and 23 mL of double distilled water, sonicate, mix well, add dropwise 5 mL of 25% ammonia solution, keep stirring at 30°C for 2 h, and centrifuge at 6000 rpm for 15 min. The supernatant was discarded, and the collected precipitate was washed with ethanol and deionized water until the pH value was neutral, and the SiO-containing 2 solution of nanoparticles; FeSO with a molar ratio of 1:5 4 and K 3 Fe(CN) 6 added to the SiO-containing 2 In the solution of nanoparticles, keep stirring at room temperature for 12h, centrifuge at 6000rpm for 20min, discard the supernatant, collect the precipitate and wash it with ethanol and deionized water respectively to obtain the PB-SiO 2 solution of nanoparticles; to the containing PB-SiO 2 Add N-(β-aminoethyl)-aminopropyltriethoxysilane (APTES) solution to the solution of nanopart...

Embodiment 2

[0063] Example 2 Preparation of Au-PB-SiO 2 composite nanoparticles

[0064] Add 2 mL of tetraethyl orthosilicate (TEOS) into a mixed solution of 10 mL of ethanol and 23 mL of double distilled water, sonicate, mix well, add dropwise 5 mL of 25% ammonia solution, keep stirring at 30°C for 2 h, and centrifuge at 6000 rpm for 15 min. The supernatant was discarded, and the collected precipitate was washed with ethanol and deionized water until the pH value was neutral, and the SiO-containing 2 solution of nanoparticles; FeSO with a molar ratio of 1:5 4 and K 3 Fe(CN) 6 added to the SiO-containing 2 In the solution of nanoparticles, keep stirring at room temperature for 12h, centrifuge at 6000rpm for 20min, discard the supernatant, collect the precipitate and wash it with ethanol and deionized water, respectively, to obtain the PB-SiO containing 2 solution of nanoparticles; to the containing PB-SiO 2 Add N-(β-aminoethyl)-aminopropyltriethoxysilane (APTES) solution to the solu...

Embodiment 3

[0065] Example 3 Preparation of Au-PB-SiO 2 composite nanoparticles

[0066] Add 2 mL of tetraethyl orthosilicate (TEOS) into a mixed solution of 10 mL of ethanol and 23 mL of double distilled water, sonicate, mix well, add dropwise 5 mL of 25% ammonia solution, keep stirring at 30°C for 2 h, and centrifuge at 6000 rpm for 15 min. The supernatant was discarded, and the collected precipitate was washed with ethanol and deionized water until the pH value was neutral, and the SiO-containing 2 solution of nanoparticles; FeSO with a molar ratio of 1:5 4 and K 3 Fe(CN) 6 added to the SiO-containing 2 In the solution of nanoparticles, keep stirring at room temperature for 12h, centrifuge at 6000rpm for 20min, discard the supernatant, collect the precipitate and wash it with ethanol and deionized water respectively to obtain the PB-SiO 2 solution of nanoparticles; to the containing PB-SiO 2 Add N-(β-aminoethyl)-aminopropyltriethoxysilane (APTES) solution to the solution of nanop...

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Abstract

The invention discloses an electrochemical immunoassay method based on Au-PB-SiO2 composite nano-particles. The detection method adopts a double antibody sandwich method, allows neuron-specific enolase antibodies solid-supported on an electrode surface to perform an immunoreaction with neuron-specific enolase in a sample solution, and to combine with a co-coupled substance of Au-PB-SiO2 composite nano-particles and neuron-specific enolase antibodies. Based on the electrochemical activity of Au-PB-SiO2 composite nano-particles, the detected peak current value of a cyclic voltammetry reduction peak is positively correlated with the concentration of the neuron-specific enolase, and thus the concentration of the neuron-specific enolase in the sample to be detected can be detected. The electrochemical immunoassay method provided by the invention has a corresponding linear range of 0.25-500 ng / ml, and a lower detection limit of 0.08 ng / ml, has good specificity and high sensitivity, and has important significance for the diagnosis of small cell lung cancer (SCLC).

Description

technical field [0001] The invention relates to the field of electrochemical immunity, in particular to an Au-PB-SiO-based 2 Electrochemical Immunoassay for Composite Nanoparticles. Background technique [0002] Lung cancer is one of the most common malignant tumors in my country, and its morbidity and mortality rank first in the world, and are increasing year by year. Small-cell lung cancer (SCLC) is the most malignant type of lung cancer, accounting for 15-20% of all lung cancer cases. Tumor cells have specific biological behaviors such as low degree of differentiation, short doubling time, high division index, and early distant metastasis. They are highly sensitive to radiotherapy and chemotherapy. Most patients develop rapidly and die in a short period of time. Early diagnosis of lung cancer, formulating a reasonable treatment plan and evaluating the curative effect are particularly important. At present, the main methods used for auxiliary diagnosis of lung cancer ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/68G01N27/48B82Y40/00
Inventor 仲召阳李梦侠王东卿毅戴楠关伟
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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