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Biosensor based on surface enhanced Raman technology, and preparation method

A surface-enhanced Raman and biosensor technology, applied in nanotechnology for materials and surface science, nanotechnology for sensing, nanotechnology, etc., can solve problems such as limited sensitivity

Active Publication Date: 2020-02-04
BOZHOU CITY THE NEW HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, multiplex detection of CA19-9, MMP7, and MUC4 in serum samples from pancreatic cancer patients has been demonstrated on a gold substrate-gold nanoparticle sandwich platform with detection limits as low as 2 ng / mL
However, previous sandwich immunoassay design methods mostly used smooth macroscopic glass, gold, silver film or bimetallic film substrates, resulting in limited sensitivity.
[0005] In order to improve the sensitivity of SERS detection, Manla detection molecules are currently selected to be bonded to the base material of SERS detection, and the best effect is P-ATP molecule, the sensitivity is ng / ml-pg / ml, although the bonded P-ATP After molecules, the sensitivity of SERS detection has been improved, but the detection sensitivity and detection range width required by many detected substances are higher

Method used

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  • Biosensor based on surface enhanced Raman technology, and preparation method
  • Biosensor based on surface enhanced Raman technology, and preparation method
  • Biosensor based on surface enhanced Raman technology, and preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0020] The present embodiment provides a sandwich immunoassay biosensor based on surface-enhanced Raman technology, using the sensor to carry out SERS immunoassay to IL-6 solution and blank sample with a concentration of 1 fg / mL, the specific process is as follows:

[0021] 1. Preparation of gold nanorods

[0022] 1) Dissolve 0.1mL of 25mM HAuCl in a 20mL glass vial 4 The solution was diluted to 5mL with deionized water, and 5mL of 0.2M CTAB solution was added to the diluent to obtain solution 1;

[0023] 2) Add 0.6mL of 0.01M NaBH 4 The solution is quickly injected into solution one, NaBH 4 The solution is ready-to-use, and the mixed solution is magnetically stirred at a speed of 1200rpm, stirred for 2min, and the finally obtained seed solution is left to stand at 30°C for 30min for use;

[0024] 3) Dissolve 7.0g of CTAB and 1.234g of sodium oleate in 250mL of 50°C water, cool naturally to 30°C, then add 18ml of 4.0mM silver nitrate solution and keep warm for 1min to obtai...

Embodiment 2

[0041]The present embodiment provides a sandwich immunoassay biosensor based on surface-enhanced Raman technology, using the sensor to measure KIM1 in artificial urine, specifically as follows:

[0042] Using the structure of the sandwich immunodetection biosensor based on surface-enhanced Raman technology in Example 1, the top layer is bonded to KIM1 detection antibody, and the substrate is bonded to KIM1 capture antibody, as follows:

[0043] First, 6.0 ml of vitamin K4-labeled gold-core silver-shell nanorod solution was taken, centrifuged at 7000 rpm for 10 minutes and dispersed into 6.0 mL of deionized water. Next, 12 μL, 25% wt glutaraldehyde solution was added to 6.0 ml vitamin K4-labeled gold-core silver-shell nanorod solution, shaken gently for 1.5 h, the product was centrifuged at 6000 rpm for 10 min, and the precipitate was dispersed into 6 ml, 9 μg / mL KIM1 detection antibody solution to obtain the top structure solution, the mixed solution was stored at 4°C for 12h,...

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Abstract

The invention provides a biosensor based on a surface enhanced Raman technology, and a preparation method. The biosensor is a three-layer sandwich structure, which comprises an upper-layer structure,a middle-layer structure and a lower-layer structure; the upper-layer structure is a gold-core and silver-shell nanorod bonded with Raman detection molecules and a detection antibody; the middle-layerstructure is an antigen; and the lower-layer structure is a gold-core and silver-shell nanorod bonded with a capture antibody. According to the sandwich immunoassay biosensor based on the surface enhanced Raman technology in the invention, due to combination between the Raman detection molecules and the gold-core and silver-shell nanorod, the detection signal is greatly enhanced; the sensitivityon ultra-trace substance and single molecule detection is obviously increased; and the minimum limit can reach fg / ml.

Description

technical field [0001] The invention relates to the field of Raman scattering, in particular to a sandwich immunoassay biosensor based on surface-enhanced Raman technology. Background technique [0002] At present, the commonly used immunoassay method is enzyme-linked immunosorbent assay (ELISA). This method detects trace substances in body fluids through specific binding of antibodies and enzyme complexes, and then performs chromogenic detection. The detection limit of this method can reach pg / ml, however, substances requiring higher sensitivity cannot be detected. [0003] Therefore, in order to reliably and accurately detect ultra-trace or even single-molecule level analytes, surface-enhanced Raman (SERS) has been gradually developed and widely used in various chemical sensing, bioanalysis, biosensing and early areas of cancer diagnosis. A typical SERS immunoassay detection method is based on a sandwich immune complex capable of capturing antibodies immobilized on a sol...

Claims

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Application Information

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IPC IPC(8): G01N21/65B82Y5/00B82Y15/00B82Y30/00B82Y40/00
CPCG01N21/658B82Y5/00B82Y15/00B82Y30/00B82Y40/00
Inventor 张茂峰熊良钟熊清爵陈敏阮志燕王振
Owner BOZHOU CITY THE NEW HEALTH TECH CO LTD