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Complexation reaction amplification signal based iron ion longitudinal relaxation time sensor and construction method and purposes thereof

A technology of longitudinal relaxation time and complexation reaction, applied in instruments, scientific instruments, and nuclear magnetic resonance analysis, etc., can solve the problems of limited development, inability to meet clinical diagnosis, low sensitivity of immune sensors, etc., and achieve the effect of improving sensitivity

Pending Publication Date: 2019-11-15
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

T 1 Immunosensors can realize both biochemical analysis and immune detection, but currently T 1 The sensitivity of immunosensors is not high enough to meet the needs of clinical diagnosis, thus limiting its development

Method used

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  • Complexation reaction amplification signal based iron ion longitudinal relaxation time sensor and construction method and purposes thereof
  • Complexation reaction amplification signal based iron ion longitudinal relaxation time sensor and construction method and purposes thereof
  • Complexation reaction amplification signal based iron ion longitudinal relaxation time sensor and construction method and purposes thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Hydrogen peroxide (H 2 o 2 ) has strong oxidizing properties, Fe 2+ Ions are oxidized to Fe 3+ ions, under the action of KSCN, finally generate Fe(SCN) 3 complex, resulting in T 1 signal changes through T 1 The signal indirectly reflects the H 2 o 2 content.

[0064] Iron ion T 1 The sensor detects hydrogen peroxide (H 2 o 2 )

[0065] Add a series of different concentrations (10, 20, 40, 80, 160, 320 and 640 μM) of hydrogen peroxide (50 μL) to 100 μL of 5 mM FeCl 2 React at 37°C for 40 minutes, take 50 μL of the mixed solution, add 50 μL of water and 50 μL of 5% KSCN solution, react for 5 minutes, finally take 20 μL of the mixed solution, and use a small nuclear magnetic resonance instrument to measure T 1 signal, the result of which is figure 1 Shown, T 1 The amount of change in value varies with H 2 o 2 The concentration increases and becomes larger, and there is a good linear relationship between the two.

Embodiment 2

[0067] Glucose can produce H under the action of glucose oxidase (GOD) 2 o 2 , H 2 o 2 can cause Fe 2+ Ions are transformed into Fe 3+ ions, KSCN complexed Fe 3+ Ions generate Fe(SCN) 3 complex, resulting in T 1 signal changes through T 1 The signal indirectly reflects the content of the target substance (glucose or glucose oxidase). That is to say, many kinds of biochemical analysis indicators can be detected through this oxidation-reduction reaction.

[0068] Iron ion T 1 Sensor detects glucose

[0069] Glucose (100 μL) at a series of different concentrations (0.005, 0.01, 0.02, 0.04, 0.08, 0.16, 0.32, 0.64, 1.25, 2.5, 5, 10 and 20 mM) was added to 100 μL of glucose oxidase (0.02 mg / mL) solution React at 37°C for 1 hour, then take 50 μL of the mixture and add it to 100 μL of 5 mM FeCl 2 React at 37°C for 40 minutes, and finally take 50 μL of the above mixture, add 50 μL of water and 50 μL of 5% KSCN solution, and use a small nuclear magnetic resonance (NMR) to ...

Embodiment 3

[0073] GOD itself is an important biomarker, so while realizing GOD detection, the characteristics of GOD labeling enzyme can also be used to realize immune analysis. Because the content of the GOD-labeled secondary antibody is positively correlated with the concentration of the target, the T caused by GOD 1 The change of the signal is positively correlated with the content of the target substance in the sample, so as to realize the quantitative analysis. Therefore the T 1 The sensors enable immunoassays.

[0074] The detection of rabbit anti-human IgG was realized by immunoassay reaction.

[0075] Experimental steps:

[0076] (1) Human IgG was diluted to 5 μg / mL with coating solution (carbonate buffer, pH 9.6), and added to the wells of the ELISA plate, 100 μL / well. Placed at 37°C, reacted for 2h.

[0077] (2) At room temperature, shake off the solution in the well and pat dry. Add PBS solution containing 0.5‰ (volume) Tween-20, 150 μL / well, let stand for 1 min, shake off...

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Abstract

The invention provides a complexation reaction amplification signal based iron ion longitudinal relaxation time sensor. The sensor uses longitudinal relaxation time (T1) as a read-out signal, and theread-out signal of the longitudinal relaxation time (T1) changes with Fe2+ / Fe(SCN)3 concentration change caused by oxidation reduction reaction and complexation reaction. The invention also provides aconstruction method of the complexation reaction amplification signal based iron ion longitudinal relaxation time sensor as well as purposes of the sensor in the fields including biochemical analysis, immunization analysis, food security and environment detection.

Description

technical field [0001] The invention belongs to the technical field of biomolecular recognition, and in particular relates to an iron ion longitudinal relaxation time sensor based on complexation reaction amplification signal and its construction method and application. Background technique [0002] The magnetic relaxation time sensor uses magnetic signals for detection and does not rely on optical signals, so it can avoid the interference of complex matrices and reduce complex steps such as sample pretreatment. Magnetic relaxation time sensors have been widely concerned and applied in fields such as medical diagnosis, food safety, and environmental analysis. At present, the magnetic relaxation time sensors mainly include: (1) Transverse relaxation time (T 2 ) Immunosensor: In a uniform magnetic field, changes in the state of magnetic particles (from dispersion to aggregation, or from aggregation to dispersion) will change the uniformity of the magnetic field and cause surr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N24/08
CPCG01N24/082G01N24/08G01N24/088
Inventor 蒋兴宇董铭灵陈翊平
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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