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Electrode for biosensor for NADH measurement and manufacturing method therefor

A biosensor and electrode manufacturing technology, applied in biochemical equipment and methods, measurement devices, and microbial determination/inspection, etc., can solve the problems of complex pretreatment process, reduced selectivity, damaged electrode surface, etc., and achieve the maintenance of sensitivity. and selectivity, improved surface modification efficiency, and stable current flow

Inactive Publication Date: 2020-08-14
KOREA RES INST OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Therefore, although many studies have focused on the basic scientific application of NADH reaction, in the case of conventional absorbance method, the measurement of NADH in blood has very serious interference phenomenon, while the sensitivity is low and the selectivity is significantly reduced, and the sample consumption is very large, The preprocessing process is complicated, so it is difficult to measure easily
Moreover, the WST-1 and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) analysis methods of the related art correspond to the following method: After the substance, induce the NADH factor and color reaction to measure the absorbance at a specific wavelength, but this method requires a large number of samples, the reaction time is long, and it is not suitable for continuous and real-time monitoring due to poor decomposition ability
Also, in order to measure NADH electrochemically, after inducing an enzyme reaction or putting in a mediator of ruthenium or cyanide ions which are catalytic materials actively performing electron transport, the redox current of NADH is measured, but in this case, Since oxidation and reduction usually occur at a voltage value above 1000mV, the surface of the electrode is damaged, and it is impossible to repeat the measurement, and the decomposition ability is significantly reduced

Method used

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  • Electrode for biosensor for NADH measurement and manufacturing method therefor
  • Electrode for biosensor for NADH measurement and manufacturing method therefor
  • Electrode for biosensor for NADH measurement and manufacturing method therefor

Examples

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manufacture example

[0055] [Manufacturing example] Manufacture of a biosensor electrode for NADH measurement

[0056] An electrode suitable for a biosensor for NADH measurement was fabricated by performing the following steps.

[0057] With 10mM sulfuric acid (H 2 SO 4 ) to wash the gold (Au) electrode. Then, in order to form a self-assembled film, the electrode was immersed in 4-aminothiophenol prepared at a molar concentration of 10 mM, followed by incubation for 2 hours. Thereafter, the electrode is immersed in a phosphate buffer solution with a molar concentration of 100 mM (high concentration), and a voltage sweep is performed from 0.8 V to -0.4 V potential by cyclic voltammetry to form N-phenylquinone di imine (hereinafter referred to as "NPQD"). After immersing the electrode formed with the NPQD into a phosphate buffer with a molar concentration of 10 mM (low concentration), apply the same voltage as in the step of forming the NPQD in the 100 mM phosphate buffer to perform voltage sc...

Embodiment 1

[0060] [Example 1] 4-aminothiophenol contact angle measurement

[0061] Changes in the contact angles of the NPQDs formed on the surfaces of the electrodes according to the manufacturing examples were confirmed. Specifically, after 10 μl of distilled water was dropped on the electrode of the preparation example at room temperature and a humidity of 46%, each electrode was photographed and analyzed by using the method provided by the manufacturer through the IMAGE J software to measure and analyze the contact angle, which result in figure 1 with figure 2 shown in .

[0062] Such as figure 1 with figure 2 As shown, it can be seen that the NPQDs bound to the surface of the preparation example can be bound to the surface in a larger amount, whereby the surface of the electrode is converted to have greater hydrophilicity. In particular, as figure 2 As shown, there was little change in the contact angle between the comparative example and the case where only 4-aminothio...

Embodiment 2

[0064] [Example 2] Measuring comparative current value stability

[0065] In order to compare the stability of the current value of high-concentration phosphate buffer (100mM) and low-concentration phosphate buffer (10mM), the CH1040C series multipotentiometer device was used to perform electrochemical analysis using the protocol provided by the manufacturer. The result is as image 3 shown.

[0066] Such as image 3 As shown, the current value measured in the high-concentration phosphate buffer solution is relatively unstable, but the current value is very stable in the low-concentration phosphate buffer solution of 10 mM.

[0067] From the above results, it can be seen that in the production according to the production example of the present invention, the current value can be stabilized by performing the polymerization reaction by changing the steps under the conditions of high concentration and low concentration.

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Abstract

The present invention relates to an electrode for a biosensor for NADH measurement and a manufacturing method therefor. The electrode manufactured by the method according to the present invention enjoys the advantages of stabilizing current flow during an electric polymerization reaction, making the contact angle of the modified material remarkably small to increase the efficiency of surface modification, and being reusable several times. In addition, when being applied to the biosensor for NADH measurement, the electrode of the present invention maintains sensitivity and selectivity at a highlevel without interference and thus easily measures a target of interest even in blood or serum that necessarily requires a pretreatment process due to the existence of a trace amount of a material to be measured. In addition, when being applied to the biosensor for NADH measurement, the electrode can measure cell viability in a continuous manner and in real time, which leads to the application thereof to the cell toxicity assay field, and enables the measurement of cell viability in apoptotic cells lacking the mitochondrial function.

Description

technical field [0001] The present invention relates to an electrode for a biosensor and a manufacturing method thereof, in particular, to an electrode for a biosensor for measuring NADH (reduced state of NAD, reduced state of nicotinamide adenine dinucleotide) and a manufacturing method thereof. Background technique [0002] With the development of biotechnology, the requirements and demands for faster and more accurate analytical techniques are gradually increasing. Among these analytical techniques, many studies on biosensors have recently been conducted. In particular, applying electronic engineering by simulating biological functions, there is an increasing demand for biosensors, which are biological devices capable of sensing physical or chemical stimuli from the outside. These biosensors have attracted attention because of their very high selectivity or sensitivity to measurement objects. [0003] So far, various studies have been conducted on biosensors, and these ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327C12Q1/00
CPCC12Q1/008C12Q1/001G01N27/3276G01N27/3272G01N27/3275G01N27/3277
Inventor 李圭弘
Owner KOREA RES INST OF CHEM TECH
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