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Preparation method and application of L-cysteine/gold nanoparticle composite film cell sensor for allergen detection

A cell sensor and gold nanoparticle technology, which is applied in the field of allergen detection with sensitization properties, can solve the problems of no unified experimental method, low detection sensitivity, and different indicators used to achieve rapid detection, improved sensitivity and The effect of response speed and improved sensitivity

Inactive Publication Date: 2013-04-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When detecting degranulation of mast cells in vitro, the commonly used detection components are histamine, β-hexosaminidase, and tryptase, but the detection methods of these three are backward, there is no unified experimental method, and the indicators used are different. The operation is cumbersome, the detection sensitivity is low, and it is easy to cause a large measurement error, which limits the popularization of these methods in the analysis of allergens

Method used

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  • Preparation method and application of L-cysteine/gold nanoparticle composite film cell sensor for allergen detection
  • Preparation method and application of L-cysteine/gold nanoparticle composite film cell sensor for allergen detection
  • Preparation method and application of L-cysteine/gold nanoparticle composite film cell sensor for allergen detection

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Embodiment 1

[0021] Embodiment 1 Preparation of electrochemical cell sensor

[0022] 1. Nano gold, L-cysteine ​​modified gold electrode

[0023] After soaking the gold electrode (Φ=2mm) in the Piranha solution for 15 minutes, use 0.3 μm and 0.05 μm Al 2 o 3 Polish the powder, polish and polish the electrode surface into a mirror surface, then ultrasonically clean it with 1:1 (V:V) nitric acid, absolute ethanol, and ultrapure water for 5 minutes, blow dry with nitrogen, and set aside at 4°C. 1mM HAuCl 4 Dissolve in 0.5M H 2 SO 4 In, configure 0.05mM HAuCl 4 solution, the gold electrode was placed in the HAuCl after blowing dry with nitrogen gas 4 solution, the electrolysis potential is -0.5, the electrolysis time is 100s, blown dry with nitrogen, and set aside at 4°C. Then 0.121g of L-cysteine ​​was prepared into 0.5mg mL with acetate PB buffer -1 Wash with L-cysteine ​​solution. Soak the nano-gold-modified gold electrode at 37°C for 5 hours, wash it repeatedly with ultrapure water...

Embodiment 2

[0028] Embodiment 2 Application of electrochemical cell sensor

[0029] 1. Detection of standard substances

[0030] Weigh different quality C48 / 80 standards, and prepare them with RPMI1640 culture solution to obtain final concentrations of 0.5, 1.0, 1.5, 2.0, 2.5, 3 μg mL -1 C48 / 80 solution. The collagen-cell mixture was inoculated into a 96-well plate at 100 μL per well, and 50 μL of C48 / 80 solutions of different concentrations were added to each well in sequence, 2 wells in parallel, and incubated overnight at 37°C. Take out the modified cell sensing electrode, add 10 μL of different concentrations of C48 / 80 standard stimulated cell-collagen culture solution successively, incubate at 37°C for 30 minutes, rinse with ultrapure water, insert the electrode into the electrolytic cell for measurement .

[0031] 2. Analytical conditions and methods

[0032] The analysis method is as follows: Electrochemical workstation ATUO LAB under the conditions of the initial amplitude of ...

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Abstract

The invention discloses a method for preparing an electrochemical cell sensor. A sensitinogen standard substance is quantitatively and accurately detected, and a technical basis is provided for allergen detection. The preparation method comprises the steps of modifying the nano gold and L-cysteine materials to a gold electrode through an electro-deposition and self-assembly method; taking P815 mastocyte as a sensing medium, establishing a collagen / cell 3D culture mode by utilizing a mouse tail type I collagen, and inoculating the collagen to a modified electrode so as to finish establishment of the cell sensor; and finally, applying the cell sensor to the detection of the sensitinogen standard substance C48 / 80, presenting a good linear relationship in a concentration range of n0.5-3 mu g / mL, and obtaining the detection limit of 0.13mu g / mL. According to the method, the sensitinogen standard substance is accurately, efficiently and rapidly identified, conducted and detected, the defects in the traditional detection method are overcome, and the obtained sensor is easy to produce and low in cost and has good application prospect in allergen detection.

Description

technical field [0001] The invention relates to a preparation method and application thereof based on an electrochemical L-cysteine / gold nanoparticle composite membrane cell sensor, and belongs to the technical field of detection of allergens such as C48 / 80 with sensitization properties. Background technique [0002] In recent years, with the rapid expansion of people's diet, the incidence and prevalence of food allergies are increasing, especially with the emergence of genetically modified foods, food allergies will become more diverse, complicated and serious. Therefore, the safety evaluation of food allergens has become an important topic in the field of food safety research. At present, more than 180 kinds of allergic foods have been identified, and the Food and Agriculture Organization of the United Nations has reported that more than 90% of food allergens exist in 8 major food categories: milk, eggs, fish, crustacean aquatic products, peanuts, soybeans, nuts and wheat ...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30
Inventor 孙秀兰蒋栋磊纪剑张银志
Owner JIANGNAN UNIV
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