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Phenol recognition SERS probe, preparation and application thereof, and universal ultra-sensitive immunoassay method based on SERS

A probe and phenol technology, applied in the field of biomolecular detection, can solve the problems of insufficient enhancement of Raman signal, weak spontaneous adsorption, weak Raman signal, etc.

Active Publication Date: 2020-01-07
JINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still deficiencies in SERS spectral detection. For example, most of the substrates based on simple nanostructures of gold and silver or their alloys are not enough to enhance the Raman signal due to the lack of rich "hot spot" regions; in addition, SERS substrates have relatively For random structures, even if the substrates are prepared in the same batch, the SERS enhancement factors may still have a large difference, which makes the uniformity of the SERS signal poor; Weaker spontaneous adsorption will also result in weaker Raman signals
In addition, the problem of irregular signal fluctuations during SERS detection needs to be solved urgently, and the repeatability and reliability of SERS signals need to be improved

Method used

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  • Phenol recognition SERS probe, preparation and application thereof, and universal ultra-sensitive immunoassay method based on SERS
  • Phenol recognition SERS probe, preparation and application thereof, and universal ultra-sensitive immunoassay method based on SERS
  • Phenol recognition SERS probe, preparation and application thereof, and universal ultra-sensitive immunoassay method based on SERS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] (1) Synthesis of "cabbage" structure gold microparticles

[0081]Dissolve 8 μL of 10% chloroauric acid aqueous solution in 1 mL of 1 mmol / L hydrochloric acid, add 1.6 mg of PVP, place in ice water for 10 minutes, then add 40 μL of 100 mg / mL NAAN hydrochloric acid aqueous solution, and shake for 20 minutes Leave to react at 4°C for 24 hours; centrifuge at 5000rpm for 10min, wash with NMP after centrifugation to obtain gold microparticles CLMP, then disperse in water to obtain a CLMP aqueous solution with a concentration of 100 / mL, store at 4°C for use. The morphology of the prepared "cabbage" structure gold microparticles CLMP is as follows: figure 1 shown.

[0082] (2) Synthesis of Raman molecules that specifically recognize phenol

[0083] Dissolve 2.82mmol (700mg) DTDBA in 40mL of HCl with a concentration of 1mol / L, cool in an ice water (0°C) bath, then add 2mL of sodium nitrite aqueous solution with a concentration of 3.1mmol / mL while stirring, and place in an ice ...

Embodiment 2

[0088] A SERS detection method for phenol.

[0089] Add 10 μL of the SERS probe aqueous solution prepared in Example 1 to 1 mL of Na 2 CO 3 In aqueous solution, the Na 2 CO 3 The concentration of phenol in the aqueous solution is 1×10 -9 mol / L, 5×10 -9 mol / L, 1×10 -8 mol / L, 5×10 -8 mol / L, 1×10 -7 mol / L, 5×10 -7 mol / L, 1×10 -6 mol / L, 1×10 -5 mol / L, 1×10 -3 mol / L, the Na 2 CO 3 The mass concentration of the aqueous solution was 5%, and then both were incubated at 4°C for 15 min, the probes were collected and washed with water, and the precipitate obtained was then dispersed in water (wherein the content of SERS probes was 100 / mL) to obtain the sample to be tested. A drop of the sample to be tested was drawn and dried at room temperature for 2 h before SERS measurement.

[0090] DTDBD-CLMP and Na 2 CO 3 After incubation with phenol (concentration: 1mmol / L) in aqueous solution, DTDBD reacts with phenol to generate azobenzene, at 1141cm -1 、1391cm -1 and 1438cm -...

Embodiment 3

[0100] A kind of SERS detection method of ALP

[0101] To 950 μL of Tris-HCl buffer (pH 9.8) containing 1 mmol / L PPNa, add 50 μL of different concentrations (0.1 mU / L, 0.5 mU / L, 1 mU / L, 5 mU / L, 10 mU / L , 50mU / L) ALP aqueous solution. Incubate at room temperature for 20 minutes, add 10 μL of the SERS probe aqueous solution prepared in Example 1, and incubate at room temperature for 15 minutes. After washing with water, the resulting precipitate was dispersed in water to obtain the sample to be tested (the content of SERS probes was 100 / mL), and a drop of the sample to be tested was dropped on the silicon wafer, dried at room temperature for 2 hours, and then SERS was measured. .

[0102] The reaction mechanism and detection results of this embodiment are as follows: Figure 7 , 8 , as shown in 9. 1141cm -1 The intensity of the SERS peak at this point was obviously continuously enhanced with the increase of ALP concentration (see Figure 8 ). According to the method of t...

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Abstract

The invention discloses a phenol recognition SERS probe, a preparation and an application thereof, and a universal ultra-sensitive immunoassay method based on SERS. Firstly, a phenol responsive SERS probe is prepared by reducing chloroauric acid with DTDBA, ahen, ELISA is combined with SERS, biomolecules are labeled with ALP, a substrate PPNA is hydrolyzed by ALP to generate phenol. An SERS probesignal caused by the phenol is used for realizing a technology for sensitive detection of the biomolecules. The technology can not only overcome the disadvantages of low sensitivity of the conventional enzyme linked immunosorbent assay, but also can solve the problems of enhancement and poor reproducibility of SERS detection signals. The phenol recognition SERS probe disclosed by the invention hasremarkable universality, can be widely used for immunoassay using the ALP as an enzyme label to measure a variety of biomolecules, lays a solid foundation for the development of immune technology based on SERS detection, and has a very large development space and a broad application prospect in the fields of biological and chemical detection, medical diagnosis, and the like.

Description

technical field [0001] The invention belongs to the technical field of biomolecular detection, and in particular relates to a phenol recognition SERS probe, its preparation and application, and a SERS-based general ultrasensitive immune analysis method. Background technique [0002] The traditional enzyme-linked immunosorbent assay (ELISA) uses the specific bonding characteristics between antigens and antibodies, and designs its bonding mechanism based on the "sandwich" strategy to label the second antibody molecule with an enzyme. Utilizing the catalytic performance of the enzyme to make the corresponding substrate change in color or fluorescence, it can display the presence or absence of a specific antigen or antibody, and perform quantitative analysis by measuring the corresponding optical intensity change to detect the analyte. However, limited by the sensitivity of these spectroscopic methods, the sensitivity of ELISA immunoassay needs to be improved urgently, so this t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65G01N1/38G01N1/34
CPCG01N1/34G01N1/38G01N21/658
Inventor 李楠陈浩凌査勇超牟宗霞薛巍
Owner JINAN UNIVERSITY
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