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ELISA kit and use method thereof

A kit and antibody technology, applied in the field of immunoassay, can solve the problems of large background interference and low sensitivity, and achieve the effect of high signal reproducibility, high sensitivity, and low detection limit

Pending Publication Date: 2022-04-15
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the ELISA detection kits for IL-17A are imported, and there are still problems such as low detection sensitivity and large background interference.

Method used

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  • ELISA kit and use method thereof
  • ELISA kit and use method thereof
  • ELISA kit and use method thereof

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Embodiment 1

[0045] 1. Synthesis of PS / PDA nanoparticles

[0046] Add 5 mg of dopamine to 50 mL of Tris buffer solution containing 0.2 mg / mL polystyrene nanoparticles, and after ultrasonic dispersion is uniform, the solution is placed on a stirrer and reacted for 12 hours. After finishing, the solid product is washed several times with water to obtain polystyrene / polydopamine nanoparticles (PS / PDA) nanoparticles. The particle size of the polystyrene nanoparticles is 60-70nm, and the dopamine is self-polymerized and modified on the surface of the polystyrene under an alkaline environment to obtain PS / PDA nanoparticles of 90-100nm.

[0047] 2. Enzyme-labeled complex HRP-PS / PDA-Ab 2 Synthesis

[0048] Dissolve 1.7mg of PS / PDA nanoparticles in 1mL of PBS buffer solution, mix well by ultrasonic, add 20μL of 1mg / mL HRP solution and 2.5μL of 1mg / mL Ab 2 (Antigen detection antibody to be tested, purchased from Beijing Yiqiao Shenzhou Science and Technology Co., Ltd.) solution, mixed slightly even...

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Abstract

The invention relates to the field of immunodetection, in particular to an ELISA (enzyme-linked immunosorbent assay) kit and a use method thereof. The kit comprises a to-be-detected antigen capture antibody, an elisa plate, an elisa compound, a coating solution, a washing solution, a confining solution, a developing solution and a stop solution. Compared with a traditional ELISA method, the kit has the advantages that a polystyrene / polydopamine covalent coupling detection antibody and an enzyme-labeled compound of horse radish peroxidase are adopted to replace a horse radish peroxidase labeled detection antibody, so that a response signal is amplified, and the kit has the characteristics of high sensitivity, low background interference and the like. The synthesis method of the enzyme-linked compound is simple in step, low in cost and mild in condition, the enzyme-linked compound is very stable, and detection under different pH conditions can be realized.

Description

technical field [0001] The invention relates to the field of immunoassay, in particular to an ELISA kit and a use method. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) is a technique designed to detect and quantify (bio)molecules in liquid samples. It is a powerful tool for clinical diagnosis, food safety and environmental monitoring. However, the main limitation of traditional ELISA is its low sensitivity, which cannot meet the needs of analysis and detection of target substances in complex (biological) samples. At present, increasing the number of enzyme molecules in the enzyme-labeled complex is an effective means to improve its sensitivity. [0003] Nanoparticles, such as metal and metal oxide nanoparticles, can be used as carriers for antibodies and enzyme molecules due to their large specific surface area, high loading capacity, easy synthesis, good chemical stability, and easy biocombination. Due to their high density, metals and their met...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/535
Inventor 向娟张雪玉阳明辉陈翔朱武
Owner CENT SOUTH UNIV