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A method for evaluating the activity of antioxidant polypeptides using nanomaterial cell sensors

A technology of cell sensors and antioxidant peptides, which is applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of incomplete explanation of the effect of antioxidant substances, long cycle, unfavorable effective evaluation, etc., and shorten the test. The effect of cycle, stable vitality and fast detection speed

Active Publication Date: 2019-08-30
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is simple to operate, a single in vitro experiment cannot fully explain the role of antioxidant substances in cells and organisms; and the method of using animal experiments to detect antioxidant activity is too long to achieve effective evaluation in a short period of time.

Method used

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  • A method for evaluating the activity of antioxidant polypeptides using nanomaterial cell sensors
  • A method for evaluating the activity of antioxidant polypeptides using nanomaterial cell sensors
  • A method for evaluating the activity of antioxidant polypeptides using nanomaterial cell sensors

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Experimental program
Comparison scheme
Effect test

Embodiment 1-1

[0050] Such as figure 1 As shown, the construction of the nanomaterial cell sensor includes the following steps:

[0051] 1), electroplating of nano-platinum particles: at first the gold disc electrode (i.e. bare gold electrode) is coated with Al 2 o 3 Powder grinding and polishing, 100KHz ultrasound for 5 minutes, cleaning with ultrapure water, then connecting the electrode to the electrochemical workstation, and measuring the current value with differential pulse voltammetry, the results are as follows figure 2 shown in a; then the bare gold electrode was placed in 10mM H 2 PtCl 6 In the solution, under the conditions of voltage -0.2V and scan rate of 100mV / s, electroplating was performed for 300s, and the platinum-plated bare gold electrode was scanned by SEM, such as image 3 As shown, then connect the platinum-plated electrode to the electrochemical workstation, and measure the current value by differential pulse voltammetry, the result is as follows figure 2 as sh...

Embodiment 1-2

[0070] The construction of the nanomaterial cell sensor includes the following steps:

[0071] 1), the electroplating of nano-platinum particles: first gold disc electrode (i.e. bare gold electrode) on Al 2 o 3 Grinding and polishing in the powder, cleaning with ultrapure water after 100KHz ultrasound for 5min; then place the bare gold electrode in 8mM H 2 PtCl 6 In the solution, electroplating for 250s under the conditions of voltage -0.3V and scan rate 150mV / s;

[0072] 2) Modification of nano-silver wires: Dissolve nano-silver wires in deionized water, then drop them onto platinum-plated electrodes, and irradiate them with fluorescent lamps for 10 minutes to fix nano-silver wires on the electrode surface to obtain nano-modified electrodes;

[0073] 3) Preparation of the gel: add sodium alginate to the DMEM culture solution with a concentration of 1×, the addition amount is 0.02g / mL, and ultrasonically treat at 100KHz for 4min, so that the sodium alginate is evenly distri...

Embodiment 1-3

[0087] The construction of the nanomaterial cell sensor includes the following steps:

[0088] 1), the electroplating of nano-platinum particles: first gold disc electrode (i.e. bare gold electrode) on Al 2 o 3 Grinding and polishing in the powder, cleaning with ultrapure water after 100KHz ultrasound for 5min; then place the bare gold electrode in 8mM H 2 PtCl 6 In the solution, electroplating for 300s at a voltage of -0.1V and a scan rate of 200mV / s;

[0089] 2) Modification of nano-silver wires: Dissolve nano-silver wires in deionized water, then drop them onto platinum-plated electrodes, and irradiate them with fluorescent lamps for 15 minutes to fix nano-silver wires on the electrode surface to obtain nano-modified electrodes;

[0090] 3) Preparation of the gel: add sodium alginate to the DMEM culture solution with a concentration of 1×, the addition amount is 0.02g / mL, and ultrasonically treat at 100KHz for 4min, so that the sodium alginate is evenly distributed in th...

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Abstract

The invention relates to a nano-material cell-based biosensor, a preparation method thereof and a new method using the nano-material cell-based biosensor to evaluate the antioxidant activity of antioxidant polypeptides and relates to the field of antioxidant capacity measuring. Colonic adenocarcinoma cells are used as the sensing elements to build a cell gel three-dimensional network structure, and the cell gel three-dimensional network structure is combined with nano-material modification electrodes to evaluate the antioxidant activity of the polypeptides. The method using the cells to simulate electrochemical signals so as to evaluate the antioxidant activity of the polypeptides is novel and convenient in detection and has high practicality in the comprehensive evaluation of the cell protection effect of antioxidant substances.

Description

technical field [0001] The invention relates to the field of antioxidant capacity measurement, in particular to a new method for evaluating the antioxidant activity of polypeptides based on cell sensors. Colon adenocarcinoma cells were used as sensing elements to construct a three-dimensional network structure of cell gel, and combined with nanomaterials to modify electrodes, the antioxidant activity of polypeptides was evaluated. Background technique [0002] Free radicals are active molecules produced by organisms in normal metabolism. They are essential signaling molecules in the metabolic process of organisms, but excessive free radicals will affect the normal physiological functions of organisms. Studies have shown that the aging and disease of organisms are related to the oxidative damage of free radicals. In a normal metabolic environment, free radicals in organisms can be eliminated through antioxidant enzyme system and non-enzymatic antioxidant mediators, thereby r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/00
Inventor 张万刚邢路娟葛庆丰
Owner NANJING AGRICULTURAL UNIVERSITY
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