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SERS sensor of sandwich structure for detecting tyrosinase, and preparation and detection method thereof

A tyrosinase and sensor technology, applied in the field of surface-enhanced Raman analysis and detection, can solve the problems of insufficient detection sensitivity, insufficient sensitivity, cumbersome sample processing, etc., achieve good biocompatibility, high sensitivity, avoid mechanical damage effect

Active Publication Date: 2019-07-09
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the detection sensitivity of these methods is not high enough, the specificity is low, and the chromatography and fluorescence analysis are limited by the requirements of tedious sample handling, complicated probe synthesis or expensive instruments.
Chinese Patent No. (CN 10285423313 B) discloses a tyrosinase biosensor based on modified electrodes and its preparation method, but this method is susceptible to environmental interference, and the detection reproducibility is poor
Chinese Patent No. (CN 109239255 A) discloses a method for assaying tyrosinase and its inhibitors based on the chitosan-platinum nanoparticle catalytic chromogenic system. The method detects tyrosinase through color changes. But the sensitivity is not high enough

Method used

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  • SERS sensor of sandwich structure for detecting tyrosinase, and preparation and detection method thereof
  • SERS sensor of sandwich structure for detecting tyrosinase, and preparation and detection method thereof
  • SERS sensor of sandwich structure for detecting tyrosinase, and preparation and detection method thereof

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Embodiment 1

[0042] Preparation of a sandwich-structured SERS sensor for sensitive detection of tyrosinase

[0043] (1) Preparation of nano-gold core

[0044] Take 100ml of a chloroauric acid solution with a mass fraction of 0.01% and heat it to boiling under magnetic stirring; then take 1-2ml of a 1% sodium citrate solution, add it to the boiling chloroauric acid solution, and wait for boiling for 10-20min , the reaction is completed, cooled to room temperature, and made into nano-gold sol;

[0045] (2) Au 4MB Preparation of @Au colloid

[0046] Take 10mL nano-gold colloid solution, add 5-15μL ethanol solution of 4-mercaptobenzonitrile with a concentration of 0.1mol / L as the Raman signal molecule, stir and mix well and then centrifuge, the purpose is to wash away the unadsorbed on the nano-gold colloid The solution of 4-mercaptobenzonitrile was resuspended in 10mL ultrapure water to obtain Au@4MB colloid; add 100-250μL to 10mL of the prepared Au@4MB colloid with a concentration of 6*10...

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Abstract

The invention discloses a SERS sensor of a sandwich structure for detecting tyrosinase, and a preparation and detection method thereof. The SERS sensor utilizes the specific recognition principle of the tyrosinase antigen antibody, and a solid phase antibody-to-be-detected antigen-labelled antibody sandwich structure is formed through the combination of the solid-phase antibody on a substrate andthe labelled antibody with the antigen through the adsorption of the magnetic substrate and the enhancement effect of the surface-enhanced Raman probe, the content of the tyrosinase is analyzed by recognizing a labelled molecular SERS signal, and then a surface-enhanced Raman analysis method for detecting the tyrosinase is established. The detection method disclosed by the invention has the advantages of being quick, sensitive, simple and less in sample consumption. Furthermore, the sensor for detecting the tyrosinase has good linearity, the detection limit is low, and can reach 1ng / mL. And meanwhile, the adding standard recovery rate of the tyrosinase in the human serum sample is in the range of 96.0%-136.0%. Therefore, the SERS sensor is expected to be an effective method for clinicallydetecting tyrosinase.

Description

technical field [0001] The invention belongs to the technical field of surface-enhanced Raman analysis and detection technology, and in particular relates to a SERS sensor with a sandwich structure for detecting tyrosinase and a preparation and detection method thereof. Background technique [0002] Tyrosinase (TYR) is a copper-containing oxidase that converts monophenols or catechols into the corresponding o-quinones, and is the rate-limiting enzyme in the biosynthesis of melanin from tyrosine. Tyrosinase is also a cytoplasmic melanocyte differentiation protein, which initiates the formation of melanin by mediating the oxidation of tyrosine to some quinones, and TYR is a potential biomarker for detecting melanoma, because its expression level correlates with malignant level closely related. In addition, abnormal levels of tyrosinase may lead to serious skin diseases such as vitiligo and neurological syndromes such as Parkinson's disease. Therefore, the determination of ty...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 陆德婵先丽卢玉栋翁婷炜沈慧英吴阳朱兰瑾陈玉婷
Owner FUJIAN NORMAL UNIV
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