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Up-conversion fluorescent/magnetic nanoparticles-based immune-chromatographic test paper and making method thereof

The technology of immunochromatographic test paper and magnetic nanoparticle is applied in the field of immunochromatographic test paper and its preparation, and can solve the problems of difficulty in quantifying detection results, low detection sensitivity, and difficulty in recording and storage.

Active Publication Date: 2014-10-15
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional method of detection by colorimetric interpretation has limitations, low detection sensitivity, difficult to quantify the detection results, and difficult to record and save

Method used

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  • Up-conversion fluorescent/magnetic nanoparticles-based immune-chromatographic test paper and making method thereof
  • Up-conversion fluorescent/magnetic nanoparticles-based immune-chromatographic test paper and making method thereof
  • Up-conversion fluorescent/magnetic nanoparticles-based immune-chromatographic test paper and making method thereof

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preparation example Construction

[0044] In one embodiment of the present invention, the preparation method of described immunochromatography test strip, comprises the following steps:

[0045] Step 1) Synthesize up-converting fluorescent / magnetic nanoparticles, and transfer them from the oil phase to the aqueous solution through ligand replacement, and the surface is bisphosphoric acid PEG with maleimide groups;

[0046] Step 2) preparing a conjugate of upconversion fluorescent / magnetic nanoparticles and polyclonal / monoclonal antibodies of the analyte;

[0047] Step 3) Preparation of the sample pad and the binding-release pad, respectively soaking the sample pad and the binding-release pad in different prefabricated solutions, and fully drying them in an oven at 37°C;

[0048] Step 4) Spotting and streaking, load the up-converting fluorescent / magnetic nanoparticles and antibody markers obtained in step 2) on the binding release pad treated in step 3); respectively mark the cellulose membrane as the detection ...

Embodiment 1

[0057] Example 1 Preparation of up-converting fluorescent / magnetic nanoparticles with different emission wavelengths

[0058] 1) Green-emitting upconversion fluorescent / magnetic nanoparticles NaGdF 4 : Preparation of Yb, Er

[0059] Add 0.183 g of gadolinium chloride hexahydrate, 0.048 g of ytterbium chloride hexahydrate, and 0.005 g of erbium chloride hexahydrate into 30 mL of a mixed solvent of oleic acid and octadecene. Dehydration (100° C.) was carried out under vacuum for 2 hours, and then the reaction solution was cooled to room temperature. Sodium hydroxide and ammonium fluoride methanol solution were added to the above reaction solution. The reaction was stopped after heating to 300° C. for 1 h under a nitrogen atmosphere. After the reaction solution was cooled to room temperature, NaGdF was precipitated with ethanol 4 : Yb, Er nanoparticles, and washed three times with ethanol, centrifuged to obtain green light emitting nanoparticles, as image 3 As shown in (A),...

Embodiment 2

[0064] Example 2: Establishment of an immunochromatographic test paper method for upconversion fluorescent / magnetic nanoparticle labeling of small molecules based on a competitive method

[0065] Taking the pesticide metabolite methyl paraoxon as an example, the steps of the method include

[0066] 1) Preparation of nanoparticles and antibody conjugates

[0067] Green-emitting NaGdF in the aqueous phase 4 : Yb, Er nanoparticles were coupled with methyl paraoxon antibody by reacting maleimide with sulfhydryl groups. First, add 0.5% to 1% tris(2-carboxyethyl)phosphine (TCEP) to the polyclonal antibody solution containing 1 mg / mL methylparaoxon dispersed in 10×PBS buffer, and react at room temperature for 30 minutes. Byproducts and unreacted TCEP were removed and transferred to Tris buffer. Then disperse the nanoparticles in Tris buffer, and add it to the above antibody solution in a certain proportion, react at room temperature for 30 minutes, ultrafilter, replace the buffer ...

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Abstract

The invention relates to an up-conversion fluorescent / magnetic nanomaterial-labeled immune-chromatographic test paper and a making method thereof. The immune-chromatographic test paper is constructed on the basis of optimizing the structure and all composition parts of a test strip through combining the fluorescent and magnetic characteristics of an up-conversion fluorescent / magnetic material with an immune-chromatographic technique. The immune-chromatographic test paper comprises a support material and an upper layer, and the upper layer comprises a sample pad, a binding and releasing pad, a cellulose membrane and a water absorbing material, wherein the cellulose membrane has a detection line and a quality control line, the detection line is coated with an antigen or antibody of an object to be measured, the control line is coated with a secondary antibody, and the binding and the releasing pad is loaded with an up-conversion fluorescent / magnetic nanomaterial-labeled antibody which can be bond to the object to be measured. The invention realizes the fluorescent quantitative and potentially magnetic quantitative detection of analytes in the fields of drug residual and the like based on the up-conversion fluorescent / magnetic nanomaterial-labeled immune-chromatographic technique, and can significantly improve the detection sensitivity of a traditional colloidal gold-labeled immune-chromatographic analysis method. The above test strip has the advantages of good specificity, high sensitivity, simple, intuitive and accurate detection, low cost, wide application range, and easy popularization and application.

Description

technical field [0001] The invention relates to an immunochromatography test paper, in particular to an immunochromatography test paper and its preparation that can realize rapid, sensitive and quantitative analysis in the fields of food safety, drug detection, etc. method. Background technique [0002] Immunochromatography (immunochromatography) technology is developed in recent years, using colored particles such as colloidal gold, colloidal selenium, etc. as markers to establish a colorimetric analysis method. Compared with traditional laboratory analysis methods, immunochromatography has the following advantages: fast, simple, does not require complicated post-processing, and is suitable for on-site analysis. Based on the above advantages, immunochromatographic methods have been widely used in the field of analysis and detection. However, the traditional method of detection by colorimetric interpretation has limitations, the detection sensitivity is low, the detection ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/533
CPCG01N21/6402G01N33/54346G01N33/558G01N33/76
Inventor 高明远刘春艳侯毅
Owner INST OF CHEM CHINESE ACAD OF SCI
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