MXene probe applied to surface enhanced Raman spectroscopy (SERS) immune test

A surface-enhanced Raman and surface Raman-enhanced technology, applied in the field of MXene probes, can solve the problems of complex and expensive preparation of gold substrates/silver substrates, and achieve the effects of cheap preparation, easy preparation, and uniform properties

Pending Publication Date: 2022-03-11
FOSHAN CHANCHENG CENT HOSPITAL CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Solid-state SERS immune chip substrates include SERS-enhanced substrates such as gold substrates and silver substrates, or non-gold and silver-modified substrates, in which gold substrates / silver substrates can form a hot spot effect with SERS detection probes to achieve high-sensitivity detection; but gold Substrate / Silver substrate preparation is complex and expensive

Method used

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  • MXene probe applied to surface enhanced Raman spectroscopy (SERS) immune test
  • MXene probe applied to surface enhanced Raman spectroscopy (SERS) immune test
  • MXene probe applied to surface enhanced Raman spectroscopy (SERS) immune test

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Embodiment 2

[0027] Example 2: see figure 2 , an embodiment provided by the present invention: an MXene probe applied to surface-enhanced Raman spectroscopy (SERS) immunoassay, preparation of MXene-based SERS detection probe, (a) MXene material adsorbs a class with strong pull Dye molecules with Mann signals, such as methylene blue, thionine, rhodamine 6G and 5,5'-dithiobis(2-nitrobenzoic acid); these molecules have very strong Raman signals and are suitable for Raman detection signal output. Mix 1 mL-1.2 mL layered MXene two-dimensional material, 300 uL-500 uL 500 mg / L dye molecules (including methylene blue, thionine, rhodamine 6G and 5,5'-dithiobis(2-nitro Benzoic acid)) and 10 uL-20 uL 20% polydiallyldimethylammonium chloride (PDDA) were mixed in a 1.5 mL EP tube, and mixed on a vertical rotary mixer for 12 h-18 h to obtain the adsorbed MXene@Dye colloidal solution of dye. Centrifuge at 8000 rpm-13300 rpm for several times, discard the supernatant, and obtain MXene@Dye precipitate....

Embodiment 3

[0028] Example 3: See image 3 , an embodiment provided by the present invention: an MXene probe applied to surface-enhanced Raman spectroscopy (SERS) immunoassay, the antibody-antigen-antibody ELISA reaction, (a) adsorption binding antibody: medium binding force Add 100 uL of 10 ug / uL conjugated antibody to each well of a 96-well ELISA plate, incubate at 4 °C for 12-18 h, discard the liquid in the well, add 100 uL of 5% BSA solution to block at 37 °C for 2 h - 4 h, wash 3-5 times with 1X PBST after blocking. Prepare an ELISA reaction plate coated with capture antibody. (b) Immune reaction: Add a certain concentration of target antigen to the positive wells of the ELISA reaction plate coated with capture antibodies, add non-target antigens with the same concentration as the target antigen to the interference wells, and add no antigen to the negative wells. Incubate at 37°C for 1 h-2 h, wash with 1X PBST for 3-5 times, add 50 uL – 200 uL of MXene-based SERS detection probe, i...

Embodiment 4

[0029] Example 4: See Figure 4 , Figure 5 and Figure 6 and Figure 7 , an embodiment provided by the present invention: an MXene probe applied to surface-enhanced Raman spectroscopy (SERS) immunoassay, the MXene probe is used for surface-enhanced Raman spectroscopy (SERS) immunoassay: in poly After the ELISA reaction is carried out on the styrene 96-well plate, the Mxene detection probe is bound to the bottom of the 96-well plate, and the signal collection head of the hand-held Raman instrument with 785 nm excitation light is aligned with the bottom of the single well, and the distance between the collection head and the single well 0.8 cm-1.5 cm, the results of the characteristic peak intensity of the MXene detection probes in the present invention combined with dye molecules (methylene blue, thionine, rhodamine 6G and 5,5'-dithiobis(2-nitrobenzoic acid) Read, the excitation power is 10-50 mW, and the integration time is 5000 ms-15000 ms. For example, read the result of...

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Abstract

The invention discloses an MXene probe applied to a surface enhanced Raman spectroscopy (SERS) immune test. A technical method of the MXene probe comprises the following steps: (1) synthesizing an MXene two-dimensional material based on V2CTx; and (2) synthesizing an MXene (at) Thi (at) AuNPs two-dimensional plane Raman probe molecule, and modifying a recognition antibody (BindingAntibody) to obtain the MXene probe which can be applied to a surface enhanced Raman spectroscopy (SERS) immune test. And (3) modifying the antibody on the polystyrene ELISA plate to capture the target protein, and adsorbing the MXene probe specific recognition target on the ELISA plate. And (4) carrying out surface enhanced Raman spectroscopy (SERS) immunodetection by adopting a handheld Raman instrument. The MXene material is easy to prepare and modify, good in biocompatibility and capable of adsorbing a large number of small molecular substance thionine (Thi) with strong Raman signals and generating obvious Raman signals, and the polystyrene ELISA reaction plate serving as an SERS immunodetection substrate has the advantages of low preparation cost, uniform property, simple modification method and the like. The handheld Raman instrument is adopted for detection, and compared with traditional large-scale confocal Raman detection and traditional ELISA detection, portability can be achieved, and detection can be achieved anytime and anywhere.

Description

technical field [0001] The invention relates to the technical field of immune analysis, in particular to an MXene probe applied to surface-enhanced Raman spectroscopy (SERS) immunoassay. Background technique [0002] Immunoassay is a reliable, easy-to-use, and inexpensive method for detecting specific antibodies or antigens in a variety of biological samples. This detection method based on the specific combination of antibody and antigen has been widely used in the fields of biochemical research and clinical diagnosis. [0003] Surface-enhanced Raman spectroscopy (SERS) has the advantages of high sensitivity, good specificity, simple operation, and multi-channel detection. Combined with immunoassay technology, it can be applied to the detection of proteins, viruses, bacteria, etc. At present, SERS immunoassay technology is usually realized by immunomagnetic beads, SERS immunochromatography test strip method and solid-phase immunoassay substrate. In research at home and abr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65G01N33/53
CPCG01N21/658G01N33/53
Inventor 曾赤佳周钦崔飞云周航刘建雷庄锡伟
Owner FOSHAN CHANCHENG CENT HOSPITAL CO LTD
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