Method of detecting biological molecules by nano enzyme immune sandwich novel technology

A technology of biomolecules and nanozymes, which is applied in the field of nanomaterials and biomedical nanotechnology, can solve the problems of complicated steps, difficult to realize automatic operation, and long time-consuming

Active Publication Date: 2018-12-11
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Traditional separation techniques mainly include processes such as precipitation and centrifugation. The steps of these purification methods are complicated, time-consuming, low yield, and contact with toxic reagents, so it is difficult to realize automatic operation

Method used

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  • Method of detecting biological molecules by nano enzyme immune sandwich novel technology

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Embodiment 1

[0051] Embodiment one detects ricin

[0052] In this embodiment, the method for detecting biomolecules by the nano-enzyme immune sandwich technology is as follows:

[0053] (1) The monoclonal antibody 6A6 of ricin was coupled to silicon dioxide-embedded iron ferric oxide magnetic particles with a particle size of 1 micron by EDC-NHS activated carboxyl method to make a capture probe;

[0054] The specific steps of the coupling are as follows: Weigh 1 mg of ferroferric oxide magnetic particles modified by silica with a particle size of 1 micron, add N-hydroxysuccinimide NHS solution with a concentration of 50 mg / mL and a concentration of 50 mg / mL 50 μL of each mL of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDC solution, incubate at room temperature for 30 minutes, wash with deionized water to remove excess NHS / EDC; add 1 mL , sodium acetate solution with a pH of 6.0 and a concentration of 50mmol / L, and 100 μg of anti-ricin monoclonal antibody 6A6, mixed well,...

Embodiment 2

[0063] Example 2 Detection of tumor marker alpha-fetoprotein AFP

[0064] In this embodiment, the method for detecting biomolecules by the nano-enzyme immune sandwich technology is as follows:

[0065] (1) The monoclonal antibody Ab-1 of alpha-fetoprotein AFP is coupled to dextran-modified particle diameters on iron ferric oxide magnetic particles of 2 microns by periodic acid oxidation method to make a capture probe;

[0066] The specific steps of coupling are as follows: Weigh 1 mg of dextran-modified ferroferric oxide magnetic particles with a particle size of 2 microns and dissolve them in 1 mL of freshly prepared NaIO with a concentration of 0.05 mol / L. 4 The solution was stirred at room temperature for 20 minutes in the dark; the supernatant was removed by magnetic adsorption, and washed three times with sodium acetate buffer solution with a concentration of 1 mmol / L and a pH of 4.4; salt buffer solution (such as buffer solution prepared with sodium carbonate and sodium...

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Abstract

The invention discloses a method of detecting biological molecules by a nano enzyme immune sandwich novel technology. The method comprises the following steps: (1) coupling a first monoclonal antibodyto first magnetic nano enzyme particles to obtain a capture probe; (2) coupling a second monoclonal antibody to second magnetic nano enzyme particles to obtain a detection probe; (3) incubating a to-be-detected sample and the capture probe; (4) obtaining a capture probe-antigen compound from a solution in the step (3); (5) resuspending the capture probe-antigen compound, adding the detection probe, and carrying out incubation; (6) obtaining a capture probe-antigen-detection probe compound from the solution in the step (5); (7) adding a peroxide and a catalytic primer to develop color, and detecting the light absorbance value of the solution through an enzyme-linked instrument; and (8) measuring biological molecules of known concentration and drawing a standard curve to obtain the antigencontent in the to-be-detected biological sample. The method is simple and rapid in operating step, environment-friendly and suitable for detecting various chemical or biological molecules.

Description

technical field [0001] The invention relates to the fields of nanomaterials and biomedical nanotechnology. More specifically, it relates to a method for detecting biomolecules using a nano-enzyme immune sandwich technology. Background technique [0002] With the rapid development of nanotechnology, nanomaterials, especially magnetic nanoparticles, have aroused great research interest in the field of biomedicine. Magnetic nanoparticles are a class of intelligent nanomaterials, which not only have the unique properties of nanomaterials, such as small particle size, large specific surface area, high coupling capacity, but also have magnetic responsiveness and superparamagnetism, and can be used under a constant magnetic field. Gathering and positioning, absorbing electromagnetic waves under an alternating magnetic field to generate heat. Using these properties, magnetic nanoparticles are used in biomarking and separation, nuclear magnetic resonance imaging, tissue repair, dru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/5434G01N33/54346G01N33/5436
Inventor 杜鸿雁段德民宋歌阎锡蕴丰蕾董颖魏春明高艳梅于忠山
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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