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Method for applying magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection

A nanoparticle and biodetection technology, applied in the field of nanomaterials and biomedical nanotechnology, can solve the problems of reduced enzyme activity and limited application, and achieve the effects of improving enzyme activity, increasing stability and biocompatibility

Inactive Publication Date: 2010-03-17
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But due to γ-Fe 2 o 3 The lack of ferrous iron in magnetic nanoparticles greatly reduces the activity of its own enzymes, which severely limits its application as a mimetic enzyme.

Method used

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  • Method for applying magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection
  • Method for applying magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection
  • Method for applying magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection

Examples

Experimental program
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Effect test

Embodiment

[0028] Example: Magnetic γ-Fe 2 o 3 The method and effect of DMSA nanoparticle imitation enzyme used in enzyme-linked immunological biological detection (ELISA)

[0029] Material Reagent: γ-Fe with surface modified dimercaptosuccinic acid 2 o 3 Magnetic nanoparticles (γ-Fe 2 o 3 / DMSA), synthesized by the laboratory according to the literature method. 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, EDC), N-hydroxysuccinimide ( N-hydroxysuccinimide, NHS), 3,3,5,5-tetramethylbenzidine (3,3,5,5-Tetramethylbenzidine, TMB) were purchased from Sigma-Aldrich Inc (USA). Goat anti-human IgG (GAHIgG), human IgG, and bovine serum albumin (BSA) were purchased from Nanjing Kaiji Biotechnology Development Co., Ltd. MES (2-(N-morpholino) ethanesulfonic acid) buffer solution at pH=6.0, PBS buffer solution (0.01M) at pH=7.4, carbonate buffer solution (0.05M) at pH=9.6, PBS / T (PBS buffer containing 0.1% Tween-20)...

Embodiment approach

[0032] 1) γ-Fe 2 o 3 / DMSA magnetic nanoparticle surface-labeled antibody. 0.2mg γ-Fe 2 o 3 / DMSA magnetic nanoparticles, dissolved in 1ml of MES, sonicated for 15min, added 10mg / ml EDC 24μl, 10mg / ml NHS 35μl, shaking reaction for 15min; after magnetic separation, MES washed 3 times, fixed solution to 1mL with MES, and Add 20 μg of goat anti-human IgG, shake and react at room temperature for 2 hours; then wash twice with PBS containing 1% BSA magnetic separation, then add 1 mL PBS containing 1% BSA for blocking for 2 hours; finally, wash twice with PBS magnetic separation and determine Dissolve to 0.2mL, store in refrigerator at 4°C, record the sample as Fe 2 o 3 -GAHIgG.

[0033] 2) ELISA detection. The polystyrene 96-well plate was irradiated with an ultraviolet lamp (UV-CJ 8F 15W) for 80 minutes on a clean bench (10 cm away), and then stored at room temperature for use. The human IgG was diluted to a concentration of 10, 5, and 2.5 μg / ml, and the coating solution wa...

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Abstract

The invention relates to a magnetic gamma-Fe2O3 nano-particle mimetic enzyme and provides a method for applying a magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection. The magnetic gamma-Fe2O3 nano-particle mimetic enzyme replaces a horse radish peroxidase (HRP) to be used in biological detection. The method comprises the following steps: coupling carboxyl groups on the surface of magnetic gamma-Fe2O3 nano-particles and a specific molecular probe to construct a specific nano probe; combining the nano probe with the specificity of the corresponding target molecules to be detected; developing by using a developer containing peroxides and hydrogen-supply zymolytes; and measuring the absorbency or carrying out the micro-observation so as to realize the qualitative and quantitative detection on target molecules.

Description

technical field [0001] The present invention relates to magnetic gamma-Fe 2 o 3 The nanoparticle simulates enzyme and provides a method for its application in biological detection, belonging to the fields of nanomaterials and biomedical nanotechnology. Background technique [0002] Magnetic iron oxide nanoparticles (Fe 3 o 4 , γ-Fe 2 o 3 ) because of its rich magnetic properties and good biocompatibility, it is widely used in biomedical fields such as magnetic resonance imaging contrast agents, magnetic targeting drug carriers, separation of cells and biomolecules, biosensing and detection, and magnetic induction tumor hyperthermia. Wide range of applications. In addition, in terms of industrial wastewater treatment, magnetic iron oxide nanoparticles are used for the removal of toxic substances such as phenol due to their high activity of hydroxylation of organic substances (the toxicity of organic substances such as phenol is reduced after hydroxylation). In recent y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/78
Inventor 张宇杨婷王春雨赵寅岗黄超彭芳芳马明顾宁
Owner SOUTHEAST UNIV
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