A black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatography test strip for detecting small-molecule substances and a preparation method thereof

A technology of gold nanoparticles and composites, applied in measuring devices, analytical materials, biological testing, etc., to achieve good accuracy and high sensitivity

Inactive Publication Date: 2019-06-18
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, photothermal quantitative immunochromatographic test strips us...

Method used

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  • A black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatography test strip for detecting small-molecule substances and a preparation method thereof
  • A black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatography test strip for detecting small-molecule substances and a preparation method thereof
  • A black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatography test strip for detecting small-molecule substances and a preparation method thereof

Examples

Experimental program
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Embodiment l

[0041] Embodiment 1 (preparation embodiment)

[0042] 1. Purification of enrofloxacin polyclonal antibody (ENR-Ab)

[0043] The present invention adopts the immunoaffinity chromatography column to purify the antiserum, and uses Protein A-Sepharose 4B as filler. The specific steps of purifying enrofloxacin antiserum are as follows:

[0044] The buffer used was sonicated for 20 min in advance to drive away air bubbles.

[0045] (1) Equilibrate the column: flush the pipeline with pH 7.4 phosphate buffer (Binding buffer) at a flow rate of 6.5 mL / min for 2 min. After loading the column, equilibrate the column with pH 7.4 phosphate buffer (Binding buffer) at a flow rate of 1 mL / min until the baseline level.

[0046] (2) Sample loading: After the baseline level, dilute the enrofloxacin antiserum with an equal volume of pH 7.4 phosphate buffer (Binding buffer) (1:1, V / V) and put it on the column at a flow rate of 0.5mL / min. After loading the sample, flush the column with pH7.4 B...

Embodiment 2

[0056] Embodiment 2 (preparation embodiment)

[0057] Assembly and preparation method of black phosphorus-gold nanoparticle complex photothermal quantitative immunochromatography test strip

[0058] 1. Test strip assembly:

[0059] The test strip of the present invention is composed as follows: sample pad 1, binding pad 2, nitrocellulose membrane 3, water-absorbing pad 4 and PVC plate 7, on the PVC plate 7, sample pad 1, binding pad 2, Nitrocellulose membrane 3, absorbent pad 4; said nitrocellulose membrane 3 is respectively coated with detection line 5 composed of enrofloxacin antigen and quality control line 6 composed of goat anti-rabbit secondary antibody.

[0060] 2. Preparation of black phosphorus-gold nanoparticles (BP-Au):

[0061] Such as figure 2 As shown, take 5mL of 200μg / mL black scale nanosheets (BPNSs) and centrifuge at 14000rpm for 15min, discard the supernatant, dissolve the precipitate in 20mL of 0.3mmol / L sodium citrate aqueous solution, add 250μL of 10m...

Embodiment 3

[0071] Embodiment 3 (application embodiment)

[0072] In order to verify the advantages of photothermal signal output, 3 μL of 6.25, 12.5, 25, 50, 100 and 200 μg / mL black phosphorus nanosheets and black phosphorus-gold nanocomposite solutions were dropped on the nitrocellulose membrane, and dried with 808nm The laser was irradiated for 30 s and the temperature was recorded with an infrared camera or an infrared thermometer to evaluate the optical image, photothermal image and temperature trend of black phosphorus nanosheets and black phosphorus-gold nanoparticle composites ( image 3 ). Optical imaging cannot see small differences between BP and BP-Au at the same concentration, and only when the concentration is increased to 50 μg / mL, the optical camera can capture the signal ( image 3 a). However, thermal imaging cameras convert grayscale images into color images through thermal signals, which can more clearly distinguish changes in material concentration on the image ( ...

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Abstract

The invention discloses a black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatography test strip for detecting small-molecule substances and a preparation method thereof. The method comprises the following steps: 1) reducing chloroauric acid to gold nanoparticles on the surface of black phosphorus; 2) adsorbing an antibody of a substance to be tested on the surfaceof the black phosphorus-gold nanoparticle compound to prepare a photothermal probe;3) pasting a sample pad, a combination pad, a nitrocellulose membrane and a water absorbent pad on a PVC back plate in sequence; 4)coating detection lines formed by an antigen of the substance to be tested and quality control lines formed by goat anti-rabbit or rat antibodies on the nitrocellulose membrane; 5)mixingthe photothermal probe and a sample to be tested and then, dropping the mixture onto the sample pad, and through the capillary action, conducting chromatography on the water absorbent pad, the antigen of the substance to be tested and the substance to be tested competitively combining the photothermal probe; and 6) carrying out laser radiation with 808 nm laser and capturing temperature with an infrared thermometer. The test strip has the advantages of high specificity, good sensitivity and simple operation and can realize quantitative detection.

Description

technical field [0001] The invention relates to a rapid detection technology of photothermal quantitative immunochromatography for small molecular substances to be tested, in particular to a black phosphorus-gold nanoparticle complex photothermal quantitative immunochromatographic test strip for detecting small molecular substances and its preparation method , belongs to the technical field of immunochromatography detection. Background technique [0002] Photothermal conversion reagents are a class of materials or compounds that can effectively convert light energy into local heat energy. Due to their good application performance, they have attracted more and more attention. They are used in energy conversion and storage, medical imaging and treatment, and transmission It has been extensively researched and applied in the fields of sensory and so on. The tumor treatment method that relies on the action of photothermal conversion reagents to generate local high temperature t...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/543
Inventor 王俊平李诗洁王硕
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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