Preparation method of immune base and antigen or antibody immunoassay method

A substrate and antibody technology, applied in the direction of measuring devices, instruments, Raman scattering, etc., can solve problems such as difficult mass production, difficult control, narrow spectral lines, etc., to avoid signal instability and non-repeatable, good biological Compatibility, the effect of narrow Raman spectral bandwidth

Active Publication Date: 2013-05-22
NINGBO UNIV
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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

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

Examples

Experimental program
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Example Embodiment

[0042] Example one:

[0043] The method for preparing an immune substrate modified with silver nanoparticles proposed in this embodiment includes 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, and then rinse the ultrasonically cleaned silicon wafer with deionized water. Dry the washed silicon wafers.

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

[0046] ①-2. Preparation of oxidant solution: Under ice bath conditions, at a volume ratio of 1:3, slowly add 30% by mass hydrogen peroxide solution to 98% by mass concentrated sulfuric acid solution to prepare An oxidizer solution is obtained. For example, take 12ml of a 98% mass percent concentrated sulfuric acid solution into a glass beaker, then pla...

Example Embodiment

[0057] Embodiment two:

[0058] This embodiment proposes a method for preparing a silver nanoparticle-modified immune substrate. The preparation method of the silver nanoparticle-modified immune substrate of this embodiment is similar to the preparation method of the silver nanoparticle-modified immune substrate 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 silver nitrate solution is 3%, and the time for the silicon wafer to be immersed in the silver nitrate solution is 2 minutes.

Example Embodiment

[0059] Embodiment three:

[0060] This embodiment proposes a method for preparing a silver nanoparticle-modified immune substrate. The method for preparing a silver nanoparticle-modified immune substrate is different from the method for preparing a silver nanoparticle-modified immune substrate given in Example 1. The process is basically the same, except that the mass percentage of hydrofluoric acid used in this embodiment is 15%, the mass percentage of the silver nitrate solution is 1%, and the time for the silicon wafer to be immersed 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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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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