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Construction of label-free nanopore sensor and application of label-free nanopore sensor in tetracycline detection

A nanopore sensor and label-free technology, which is applied in the field of nanopore sensing in analytical chemistry, can solve the problems of nanopore loss of quantitative nucleic acid molecular functions, expensive instruments, and affecting the sensing efficiency and stability of nanopore sensors.

Inactive Publication Date: 2021-02-23
TAIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these methods have been widely used in the detection of tetracycline, they all have some inevitable defects: such as the need for expensive instruments, the need for specialized personnel to operate, and the detection process takes a long time, etc.
However, when this technique was applied to environmental samples, we found that interfering ions in the environment such as Cu 2+ , Zn 2+ etc. will displace polyethyleneimine-zirconium ion complexes to form polyethyleneimine-Cu 2+ and other complexes, making polyethyleneimine-zirconium ion-modified nanopores lose the function of quantifying nucleic acid molecules
In addition, a large number of amino molecules in the environment will be adsorbed to the inner wall of the nanopore modified by polyethyleneimine-zirconium ions, which will affect the sensing efficiency and stability of the nanopore sensor.

Method used

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  • Construction of label-free nanopore sensor and application of label-free nanopore sensor in tetracycline detection
  • Construction of label-free nanopore sensor and application of label-free nanopore sensor in tetracycline detection
  • Construction of label-free nanopore sensor and application of label-free nanopore sensor in tetracycline detection

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Experimental program
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Effect test

Embodiment 1

[0013] The polyethylene phthalate (PET) film used in the experiment is a nuclear pore membrane irradiated with heavy ions by the heavy ion linear accelerator of Lanzhou Institute of Near Physics and Technology. Conical nanopores can be fabricated on heavy ion nuclear pore membranes by track etching. The ethylene phthalate (PET) film was irradiated with UV lamps (365nm and 254nm) on both sides for 1 hour each, and the irradiated samples were etched in a polytetrafluoroethylene etching tank at a temperature of 50°C. Add 5mol / L NaOH etching solution to the end of the large hole, add 1mol / L HCOOH and 1mol / L KCl etching solution to the end of the small hole, and etch for 5 hours to obtain conical nanopores. The resulting nanoporous membrane was characterized by scanning electron microscopy to determine the pore size.

Embodiment 2

[0015] Our study of surface modification of tapered nanopores. There are a large number of carboxyl groups on the surface of the tapered nanopore after chemical etching, so the non-uniform negative charges are distributed on the inner wall. We used 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) to activate the carboxyl group, and then modified the dendrimer polyamide- Amine (PAMAM) to the nanopore surface, the modification time lasted 12h. After modification, non-uniform positive charges are distributed on the inner wall of the nanopore. In the presence of graphene oxide (25 μg / mL) and 1 mM γ-cyclodextrin, the nanopore modified with PAMAM can adsorb tetracycline and DNA aptamer complex, so that the positive charge on the surface of the nanopore is partially neutralized by DNA, and the charge reduce. We can use electrochemical means to detect the transmembrane current-voltage curve (volt-ampere characteristic curve) through t...

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Abstract

The invention relates to a construction method and application of a label-free nanopore channel. The single-cone-shaped nanopore channel is prepared from a polyethylene terephthalate (PET) nuclear pore membrane by a track etching method, and carboxyl on the surface of the nanopore is activated by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) and is modified by dendritic polyamidoamine (PAMAM) (see the specification in detail). It is found through experiments that by combining the nanopore channels with graphene oxide, selective detection of tetracycline in water can be achieved, and other antibiotics do not interfere with the detection system. Therefore, the nanopore channel can be used for establishing a new method for detecting tetracycline in an environmental water sample.

Description

technical field [0001] The invention belongs to the technical field of nanopore sensing in analytical chemistry, and relates to a label-free nanopore sensor applied to the detection of tetracycline in water. Background technique [0002] Since penicillin was first discovered by humans in 1928, it has made amazing breakthroughs in the treatment of bacterial infections, and the diseases caused by bacteria have been gradually controlled. However, the excessive use of antibiotics has brought serious environmental pollution problems. Because antibiotics are difficult to metabolize in organisms, they will be discharged into natural water bodies and soil through urine and excrement, causing antibiotic residues in the environment. As one of the widely used antibiotics, tetracycline is widely used in livestock and aquaculture. Human beings consume tetracycline in low doses through drinking water and food for a long time and accumulate in the body, which will easily lead to the gener...

Claims

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Application Information

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IPC IPC(8): G01N27/00G01N27/26G01N33/18
CPCG01N27/00G01N27/26G01N33/18
Inventor 张思奇程嘉稀施伟厉凯彬
Owner TAIZHOU UNIV
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