Near-infrared fluorescent microsphere with emission peak value of greater than or equal to 1300 nm, preparation method and applications thereof

A fluorescent microsphere and near-infrared technology, applied in the field of nanomedical biotechnology detection, can solve the problems of inability to realize whole blood sample detection and unstable existence, and achieve the effect of simple biomarking process, good stability, and increased fluorescence intensity

Active Publication Date: 2020-04-10
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Some researchers have proposed to use a fluorescent probe prepared by a near-infrared fluorescent dye containing Br element with an emission peak at 1050nm for use in immunochromatographic test strips. Although the emission peak is in the near-infrared region, it is still not possible to realize the detection in whole blood. The possible reason for the detection of the sample is that the dye contains heavy atoms, which cannot exist stably in the blood

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  • Near-infrared fluorescent microsphere with emission peak value of greater than or equal to 1300 nm, preparation method and applications thereof
  • Near-infrared fluorescent microsphere with emission peak value of greater than or equal to 1300 nm, preparation method and applications thereof
  • Near-infrared fluorescent microsphere with emission peak value of greater than or equal to 1300 nm, preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1: Synthesis of near-infrared carboxypolystyrene fluorescent microspheres with an emission peak of 1300nm

[0077] The emission peak is 1300nm near-infrared carboxypolystyrene fluorescent microspheres, including carboxypolystyrene balls and near-infrared fluorescent dyes embedded in the carboxypolystyrene balls.

[0078] The particle size of the carboxylated polystyrene spheres is 300 nm.

[0079] The maximum emission wavelength of the near-infrared fluorescent dye is 1300nm, and the structure is as follows:

[0080]

[0081] The specific preparation steps of near-infrared carboxypolystyrene fluorescent microspheres are as follows:

[0082] 1. Disperse 30, 35, 40, 45 and 50 mg of near-infrared fluorescent dyes in 10 mL of tetrahydrofuran solution to form an organic phase;

[0083] 2. Take 1g of 300nm carboxypolystyrene spheres (5 in parallel) and centrifuge to remove the surfactant in the synthesis process, redissolve in 50mL ultrapure water, and fully ultr...

Embodiment 2

[0088] Example 2: Synthesis of near-infrared aminopolystyrene fluorescent microspheres with an emission peak of 1350nm

[0089] The emission peak is 1350nm near-infrared amino polystyrene fluorescent microspheres, including amino polystyrene balls and near-infrared fluorescent dyes embedded in the amino polystyrene balls.

[0090] The particle size of the aminopolystyrene spheres is 300 nm.

[0091] The maximum emission wavelength of the near-infrared fluorescent dye is 1350nm, and the structure is as follows:

[0092]

[0093] The specific preparation steps of near-infrared aminopolystyrene fluorescent microspheres are as follows:

[0094] 1. Disperse 30, 35, 40, 45 and 50 mg of near-infrared fluorescent dyes in 10 mL of chloroform solution to form an organic phase;

[0095] 2. Take 1g of 300nm aminopolystyrene spheres (5 in parallel) and centrifuge to remove the surfactant in the synthesis process, redissolve in 50mL ultrapure water, and fully ultrasonicate to form an a...

Embodiment 3

[0100] Embodiment 3: the preparation of AFP immunochromatography test strip

[0101] 1. Emission peak at 1300nm near-infrared carboxypolystyrene fluorescent microspheres coupled with AFP-Ab 1 (Jingda Bio) to prepare fluorescent probes:

[0102] 1) Take 100 mg of near-infrared carboxypolystyrene fluorescent microspheres prepared in Example 1, centrifuge, redissolve in 18 mL of BBS buffer solution with pH 7.4, and sufficiently ultrasonicate to disperse evenly to obtain a dispersion system.

[0103] 2) Add 10 mg EDC and 5 mg NHSS respectively to the dispersion system, and react at room temperature for 2 hours.

[0104] 3) After the reaction, wash by centrifugation, redissolve in 10 mL of BBS buffer solution with pH 7.4, and add 5 mg of AFP-Ab to it 1 Type monoclonal antibody, reacted at room temperature for 4 hours.

[0105] 4) After the reaction, wash by centrifugation, redissolve in 10 mL of BBS buffer solution with pH 7.4, add 100 mg of BSA to it, and react at room temperatur...

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Abstract

The invention discloses a near-infrared fluorescent microsphere with an emission peak value of greater than or equal to 1300 nm. The near-infrared fluorescent microsphere comprises a macromolecule polymer microsphere and a near-infrared fluorescent dye embedded in the macromolecule polymer microsphere, wherein the maximum emission wavelength of the near-infrared fluorescent dye is greater than orequal to 1300 nm, and the structure of the near-infrared fluorescent dye is represented by a formula I or a formula II. The invention also discloses a preparation method of the near-infrared fluorescent microsphere. According to the invention, with the application of a fluorescent probe obtained by coupling the near-infrared fluorescent microsphere and a detection antibody in an immunochromatographic test paper strip to detect a target analyte in whole blood, the penetration depth can be remarkably increased, the absorption and the scattering to emitted light by a blood sample are reduced, andthe background fluorescence signals are reduced so as to determine the target analyte in the whole blood sample.

Description

technical field [0001] The invention belongs to the technical field of nano-medicine biotechnology detection, and in particular relates to near-infrared fluorescent microspheres with an emission peak ≧1300nm and their preparation and application in immunochromatographic detection. Background technique [0002] Immunochromatography assay (ICA) or lateral flow assay (LFA) was first used to detect human chorionic gonadotropin; with the development of labeling technology, it is also widely used in medical testing, Environmental monitoring and food safety and other fields. [0003] Immunochromatography technology effectively combines chromatographic technology and antigen-antibody immune reaction technology; when the sample flows from the sample area to the adsorption area under capillary action, due to the specific interaction between the target analyte in the sample and the capture antibody , forming immune complexes during chromatography, enriched and retained on the detectio...

Claims

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Application Information

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IPC IPC(8): C09B57/00C09B67/02C09K11/02C09K11/06G01N33/558G01N33/574G01N33/577G01N33/58G01N33/76
CPCC09B67/0097C09B57/00C09K11/06C09K11/025G01N33/582G01N33/585G01N33/577G01N33/76G01N33/57476G01N33/57473G01N33/57434G01N33/57484G01N33/558C09K2211/1085C09K2211/1092C09K2211/1014
Inventor 李富友陈锐周晓波吴勇
Owner FUDAN UNIV
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