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Novel coupling process of time-resolved fluorescent microspheres and application of same

A technology of time-resolved fluorescence and microspheres, which is applied in the field of biological analysis, can solve the problems of cumbersome operation steps and low production efficiency, and achieve the effects of simplifying cumbersome procedures, low cost and saving operation time

Inactive Publication Date: 2019-12-13
JIANGSU INNOVATION BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the process of time-resolved fluorescent microsphere antibody labeling, the operation steps are cumbersome and the production efficiency is low

Method used

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  • Novel coupling process of time-resolved fluorescent microspheres and application of same
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  • Novel coupling process of time-resolved fluorescent microspheres and application of same

Examples

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Effect test

Embodiment 1

[0018] A novel time-resolved fluorescent microsphere coupling process, comprising the following steps:

[0019] (1) Activation: Add 1500ul of water, 300ul of 50mM MES activation buffer at pH 6.0, 450ul of 1% time-resolved fluorescent microsphere solution, and 150ul of 1% EDC solution into the reactor, mix well, 25 constant temperature reaction for 30 minutes;

[0020] (2) Labeling: Mix 1500ul of water, 300ul of HEPES labeling buffer with a concentration of 50M and pH6.0, and 600ul of an antibody solution with a concentration of 1.0mg / ml in the reagent bottle, and then directly add the mixed solution to the activated solution, after ultrasonic mixing, 25 ° C constant temperature reaction for 1 hour;

[0021] (3) Blocking: after the labeling reaction is completed, add 100 ul of casein blocking solution with a concentration of 5% to the solution after the labeling reaction, and react at a constant temperature of 25° C. for 3 hours;

[0022] (4) Centrifugal resuspension: After t...

Embodiment 2

[0029] A novel time-resolved fluorescent microsphere coupling process, comprising the following steps:

[0030] (1) Activation: Add 1650ul of water, 250ul of 50mM MES activation buffer at pH 6.5, 500ul of 1% time-resolved fluorescent microsphere solution, and 100 of 1% EDC solution into the reactor, mix well, 30°C constant temperature reaction for 60min;

[0031] (2) Labeling: Mix 1750ul of water, 250ul of HEPES labeling buffer with a concentration of 50mM and pH8.0, and 500ul of an antibody solution with a concentration of 1.0mg / ml in the reagent bottle, and then directly add the mixed solution to the activated solution, after ultrasonic mixing, 26 ° C constant temperature reaction for 2 hours;

[0032] (3) Blocking: After the labeling reaction is finished, add 100-150ul of 5% casein blocking solution to the solution after the labeling reaction, and react at a constant temperature of 30°C for 2h;

[0033] (4) Centrifugal resuspension: After the sealing is completed, centrif...

Embodiment 3

[0036] A novel time-resolved fluorescent microsphere coupling process, comprising the following steps:

[0037] (1) Activation: Add 1750ul of water, 250ul of 50mM MES activation buffer at pH 6.5, 500ul of 1% time-resolved fluorescent microsphere solution, and 125ul of 1% EDC solution into the reactor, mix well, 25°C constant temperature reaction for 60min;

[0038] (2) Labeling: Mix 1750ul of water, 250ul of HEPES labeling buffer with a concentration of 50mM and pH7.5, and 500ul of an antibody solution with a concentration of 1.0mg / ml in the reagent bottle, and then directly add the mixed solution to the activated solution, after ultrasonic mixing, 25 ° C constant temperature reaction for 2 hours;

[0039] (3) Blocking: after the labeling reaction, add 125 ul of casein blocking solution with a concentration of 5% to the solution after the labeling reaction, and react at a constant temperature of 25° C. for 3 hours;

[0040] (4) Centrifugal resuspension: After the sealing is ...

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Abstract

The invention provides a novel coupling process of time-resolved fluorescent microspheres. The process comprises steps of activation, labeling, confining and centrifugal resuspension. Centrifugation is not required after activation, a labeling mixed solution is directly added to an activated solution, and a final product is stored at 2-8 DEG C for later use; and the product can be used for pepsinogen I detection, pepsinogen II detection and ferritin detection. The novel antibody-labeled time-resolved fluorescent microsphere coupling process provided by the invention overcomes defects in the prior art, simplifies the tedious process, reduces operation time, improves the production efficiency, and is high in repeatability, low in cost and convenient for large-scale production and application.

Description

Technical field: [0001] The invention relates to biological analysis technology, in particular to a novel process for coupling antibody-labeled time-resolved fluorescent microspheres and the application of antibody-labeled time-resolved fluorescent microspheres. Background technique: [0002] When detecting certain antigens or specific proteins, immunolabeling techniques are usually applied. Immunolabeling technology is to label substances that are both easy to measure and highly sensitive to specific antigens or antibody proteins. [0003] Time-resolved fluorescent immunoassay is a more commonly used ultra-sensitive immunoassay method. Time-resolved fluorescent microspheres are special functional microspheres. Each microsphere can contain thousands of fluorescent molecules, which greatly improves the fluorescence labeling. efficiency. At present, in the process of antibody labeling with time-resolved fluorescent microspheres, the operation steps are cumbersome and the pro...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/533
CPCG01N21/6408G01N21/6428G01N21/6486G01N33/533
Inventor 吴红兵刘琴刘扬雷瑞斌
Owner JIANGSU INNOVATION BIOTECH CO LTD
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