Microsphere composition for chemiluminescence detection and application thereof

A chemiluminescence detection and composition technology, applied in the field of chemiluminescence, can solve problems such as narrow detection range or linear range, and achieve the effects of reducing non-specific adsorption, improving accuracy and reducing influence

Pending Publication Date: 2020-01-31
上海索昕生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the detection sensitivity of the chemiluminescence detection method can be improved to a certain extent by adopting the method described in the art, the detection range or linear range is relatively narrow

Method used

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  • Microsphere composition for chemiluminescence detection and application thereof
  • Microsphere composition for chemiluminescence detection and application thereof
  • Microsphere composition for chemiluminescence detection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Example 1: Preparation of acceptor microspheres

[0116] 1. Prepare a 25mL round-bottomed flask, add 0.1g of europium(Ⅲ) complex, 10mL of 95% ethanol, stir magnetically, and raise the temperature of the water bath to 70°C to obtain a solution of europium(Ⅲ) complex.

[0117] 2. Prepare a 100mL three-neck flask, add 10mL 95% ethanol, 10mL water and 10mL polystyrene microspheres coated with carboxydextran hydrogel with a particle size of 200nm at a concentration of 10%, stir magnetically, and heat the water bath to 70 ℃.

[0118] 3. Slowly drop the europium(III) complex solution in step 1 into the three-necked flask in step 2, react at 70°C for 2 hours, stop stirring, and cool naturally to obtain an emulsion.

[0119] 4. Centrifuge the above emulsion for 1 hour at 30,000 g, discard the supernatant after centrifugation, and then resuspend with 50% ethanol. After repeated centrifugal washing for 3 times, it was resuspended with 50 mM CB buffer solution with a pH value of ...

Embodiment 2

[0121] Example 2: Receptor microspheres coated with antibodies

[0122] 1. Measure 10 mg of receptor microspheres coated with carboxydextran hydrogel in a centrifuge tube according to the prepared amount, and centrifuge at 10,000 rpm for 60 min.

[0123] 2. Discard the supernatant, add 2 mg of Anti-PCT antibody I to the precipitate (it can be the corresponding antibody embodiment of any other analysis item (anti-cTnI antibody I and anti-PCT antibody I)), 50 μL of Tween-20 ( 50mg / mL), and a certain volume of 0.05M MES pH=6.0 was added to make the final concentration of acceptor microspheres 10mg / mL.

[0124] 3. Ultrasound to mix rapidly.

[0125] 4. Add 50 μL of NaBH to the centrifuge tube 3 CN (50mg / mL, prepared in 0.05M MES pH=6.0) was mixed evenly, and placed in a rotary mixer at 37°C for 36-48h.

[0126] 5. Blocking: add 1 mL of BSA (50 mg / mL, prepared in 0.05M MES pH=6.0), and place in a rotary mixer at 37°C for 12-16 hours.

[0127] 6. Washing: wash 3 times with 0.05M...

Embodiment 3

[0129] Example 3: Preparation of Donor Microspheres

[0130]1. Prepare a 25mL round-bottomed flask, add 0.1g of copper phthalocyanine (II) and 10mL of DMF, stir magnetically, and raise the temperature of the water bath to 70°C to obtain a copper (II) phthalocyanine solution.

[0131] 2. Prepare a 100mL three-neck flask, add 10mL 95% ethanol, 10mL water and 10mL polystyrene microspheres coated with aldehyde dextran hydrogel with a particle size of 200nm at a concentration of 10%, stir magnetically, and heat the water bath to 70°C.

[0132] 3. Slowly drop the copper phthalocyanine (II) solution in step 1 into the three-necked flask in step 2, react at 70°C for 2 hours, stop stirring, and cool naturally to obtain an emulsion.

[0133] 4. Centrifuge the above emulsion for 1 hour at 30,000 g, discard the supernatant after centrifugation, and resuspend with 50% ethanol. After repeated centrifugal washing for 3 times, it was resuspended with 50 mM CB buffer solution with a pH value...

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Abstract

The invention relates to a microsphere composition for chemiluminescence detection and application thereof in the technical field of chemiluminescence. The microsphere composition comprises donor microspheres and receptor microspheres, wherein the donor microspheres can generate active oxygen in an excited state, and the surfaces of the donor microspheres are coated with markers; the receptor microspheres can react with the active oxygen to generate detectable chemiluminescence signals, the surfaces of the receptor microspheres are coated with biomolecules, and the biomolecules can be specifically combined with target molecules to be detected; the particle size of the donor microspheres is larger than that of the receptor microspheres. According to the microsphere composition, by controlling the matrix and particle size of the luminescent microspheres and the matrix and particle size of the photosensitive microspheres, when the luminescent microsphere composition is used for immunoassay detection, the detection luminous efficiency is improved, and the good detection sensitivity is achieved.

Description

technical field [0001] The invention belongs to the technical field of chemiluminescence, and in particular relates to a microsphere composition for chemiluminescence detection and an application thereof. Background technique [0002] So far, the determination methods of trace biologically active substances in organisms have gone through various stages such as radioimmunoassay (RIA), fluorescence immunoassay (FIA), enzyme-linked immunoassay (EIA) and chemiluminescence analysis. This evolution process is mainly based on the continuous improvement of the sensitivity, accuracy, and simplicity of operation of the detection method. [0003] Chemiluminescence analysis is a detection method using light waves emitted by chemiluminescent substances. Chemiluminescent substances are used as labels in nucleic acid detection and immunodetection. For example, a certain molecule in a specific binding pair can be combined with a luminescent substance in various ways to form a luminescent ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N21/01
CPCG01N21/76G01N21/01Y02A50/30
Inventor 杨阳赵卫国刘宇卉李临
Owner 上海索昕生物科技有限公司
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