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Preparation method of immune base and antigen or antibody immunoassay method

A substrate and antibody technology, applied in measurement devices, instruments, Raman scattering, etc., can solve the problems of difficult mass production, narrow spectral lines, difficult control, etc., to avoid signal instability and non-repeatability, Raman The effect of narrow spectral bandwidth and good biocompatibility

Active Publication Date: 2015-03-18
NINGBO UNIV
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AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of antigens and antibodies mainly include: radioimmunoassay, enzyme-linked immunoassay, fluorescence immunoassay and surface-enhanced Raman scattering (SERS) immunoassay, etc. These methods have their own characteristics, for example: radioimmunoassay The detection sensitivity of analytical method and fluorescent immunoassay method is high, but the radiation and pollution of radioactive substances, the emission spectrum of traditional fluorophores are relatively wide (full width at half maximum (FWHM) is about 50-100nm), and they are easy to photodegrade, etc. Problems limit the application of radioimmunoassay and fluorescence immunoassay; enzyme-linked immunoassay has the function of high-throughput detection, but it is not suitable for ultra-sensitive detection; surface-enhanced Raman scattering (SERS) immunoassay has high sensitivity, The spectral line is narrow, suitable for high-throughput non-destructive detection analysis, but the use of SERS effect for immunodetection requires a good SERS substrate
Although SERS substrates with high precision and complex shapes can be prepared by micro-nano processing techniques such as lithography, electron beam etching, reactive ion etching, and nanoimprinting, the structure of the micro-nano processing equipment used is complex and expensive. Moreover, the production process is time-consuming, high in cost, and the prepared substrate has a small area and is not easy to mass-produce
It is relatively easy to synthesize metal (gold and silver) nanoparticles as SERS substrates by chemical methods, but the metal nanoparticles are in the solution phase. During the detection process, the nanoparticles are easy to randomly aggregate and difficult to control. The obtained SERS signal Poor stability and repeatability

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  • Preparation method of immune base and antigen or antibody immunoassay method
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Examples

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Embodiment 1

[0043] The preparation method of a silver nanoparticle-modified immune substrate proposed in this embodiment comprises the following steps:

[0044] ①-1. Take a square single-sided polished silicon wafer with a side length of 5mm, immerse it in an organic solvent, and perform ultrasonic cleaning on the silicon wafer for 30 minutes, then rinse the silicon wafer after ultrasonic cleaning with deionized water, and then Dry the rinsed wafers.

[0045] Here, silicon wafers of other shapes and sizes can also be used for single-side polished silicon wafers; acetone, alcohol or toluene can be used as organic solvents.

[0046] ①-2. Preparation of oxidant solution: under ice bath conditions, slowly add 30% mass percent hydrogen peroxide solution to 98 mass percent concentrated sulfuric acid solution at a ratio of 1:3 by volume to prepare An oxidant solution is obtained. For example, take 12ml of concentrated sulfuric acid solution with a mass percentage of 98% and pour it into a glas...

Embodiment 2

[0058] This example proposes a method for preparing an immune substrate modified by silver nanoparticles. The method for preparing an immune substrate modified by silver nanoparticles in this example is the same as the method for preparing an immune substrate modified by silver nanoparticles given in Example 1. The process is basically the same, except that the mass percentage of hydrofluoric acid used in this embodiment is 5%, the mass percentage of the silver nitrate solution is 3%, and the time for the silicon wafer to be immersed in the silver nitrate solution is 2 minutes.

Embodiment 3

[0060] This example proposes a method for preparing an immune substrate modified by silver nanoparticles. The process is basically the same, except that the mass percentage of hydrofluoric acid used in this embodiment is 15%, the mass percentage of silver nitrate solution is 1%, and the time for immersing the silicon wafer in the silver nitrate solution is 3 minutes.

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Abstract

The invention discloses an immune base preparation method and an antigen or antibody immunoassay method. According to the immune base preparation method, a silicon slice is modified by using gold or silver nanoparticles, and antibodies / antigens are modified on the surfaces of the gold or silver nanoparticles, thereby being used for capturing antigens / antibodies to be assayed. According to the antigen or antibody immunoassay method, an immune base which is prepared by using the immune base preparation method and gold or silver nanoparticle immunoprobes is used, an immune base-antigen / antibody-immunoprobe three-layer sandwich structure is formed through an antigen-antibody immune complex reaction, and the assay on the antigens / antibodies to be assayed is realized in a manner that the characteristic dactylograms of Raman markers on the surfaces of the immunoprobes are assayed by using a surface-enhanced Raman scattering effect of the gold or silver nanoparticles. The method has the advantages that the sensitivity of the immunoassay can be greatly improved and the assay on high-flux multiple antigens / antibodies can be carried out.

Description

technical field [0001] The invention relates to a Raman spectrum detection technology, in particular to a preparation method of an immune substrate and an immunological detection method of an antigen or antibody. Background technique [0002] Raman scattering is the inelastic scattering of photons, resulting from molecular vibrations or rotational energy level transitions. Raman scattering spectroscopy can be used to analyze the material composition and content changes of biological tissues and cells at the molecular level. Because Raman scattering spectroscopy is fast, non-destructive, and highly sensitive, it is widely used in the detection and analysis of life sciences, biochemical materials, and drugs. However, the Raman signal is generally weak and cannot meet the requirements of high-sensitivity detection and analysis. In 1974, Fleischmann et al. first discovered the Surface-Enhanced Raman scattering (SERS) phenomenon from the Raman scattering experiment of the mono...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/558G01N21/65
Inventor 周骏束磊马亚楠王彬彬赖魏张琪颜承恩
Owner NINGBO UNIV
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