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A method for antibody modified nano-microsphere electrophoresis flow type ELISA

A nano-microsphere and antibody modification technology is applied in the preparation of antibody-modified nano-microspheres and in the field of enzyme-linked immunosorbent assay for electrophoresis-driven NP-Ab flow detection, which can shorten the reaction time and improve the sensitivity.

Active Publication Date: 2019-04-26
BEIJING UNIV OF CHEM TECH
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Problems solved by technology

[0003] The purpose of the present invention is to solve the problem of low efficiency of sample detection in the ELISA system and further solve the problem of low sensitivity, and provide a polyelectrolyte multilayer film substrate electrophoretic flow type ELISA method, especially provide a NP-Ab electrophoretic flow type ELISA Method, to solve the problems existing in the background technology

Method used

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  • A method for antibody modified nano-microsphere electrophoresis flow type ELISA
  • A method for antibody modified nano-microsphere electrophoresis flow type ELISA
  • A method for antibody modified nano-microsphere electrophoresis flow type ELISA

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Embodiment 1

[0023] 1. Solution configuration:

[0024] a) Configuration of 50mM Tris-HCl@0.15M NaCl solution: Dissolve 1.15175g Tris-base solid in 125mL ultrapure water, take 0.84mL HCl and dissolve it in 100mL ultrapure water to obtain a 0.1M HCl solution. Add the HCl solution dropwise to the Tris-base solution to adjust the pH to 7.4, set the volume to 250 mL, weigh 2.208 g of NaCl solid and add it to the Tris-HCl.

[0025] b) Configuration of pH=7.4 concentration of 0.15M phosphate buffer (PB): weigh 10.74g Na 2 HPO 4 12H 2 Add O solid to 200mL ultrapure water, weigh 4.68g NaH 2 PO 4 2H 2 O solid was added into 200mL ultrapure water, and NaH 2 PO 4 solution added to Na 2 HPO 4 In the solution, adjust pH=7.4.

[0026] c) 0.05% Tween 20@0.03M PB configuration: Take 250 μL Tween-20 and add it to 500 mL 0.03M PB buffer.

[0027] d) 2M H 2 SO 4 Configuration: take 8mL ultrapure water and add 1mL concentrated H 2 SO 4 (98%).

[0028] 2. Preparation method of antibody-modified...

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Abstract

The invention relates to an antibody modified nanoparticle electrophoresis flowing type ELISA (enzyme linked immunosorbent assay) method and belongs to the field of biological materials. The method is characterized in that a first antibody attaches to the surface of a porous film, and sealant protein absorption is performed; antigen-first antibody reaction is performed; electrophoresis is used to drive the antibody to modified nanoparticles to perform second antibody-antigen reaction, and developing quantification is performed on an enzyme-labeled second antibody; the porous film is cleaned after the electrophoresis is used to drive the second antibody-antigen reaction, and the obtained porous film substrate is placed into the substrate solution of second antibody labeling enzyme for developing; antigen quantitative analysis is performed by detecting light absorbency. The method has the advantages that positive PDDA / PSS is used to modify a cellulose acetate membrane PEMs-CA, and carboxyl polystyrene nanoparticles are used as the carriers modified by the enzyme-labelled second antibody; PEMs modification lowers nonspecific absorption, antibody modified nanoparticles can amplify signals, electrophoresis drive force allows the second antibody to locally concentrates to the periphery of the antigen in a short time, and fast and sensitive detection is achieved.

Description

technical field [0001] The invention belongs to the technical field of biological materials, in particular to the preparation of antibody-modified nano-microspheres (NP-Ab) and an enzyme-linked immunosorbent assay (ELISA) method for electrophoresis-driven NP-Ab flow detection. Background technique [0002] Enzyme Linked Immunosorbent Assay (ELISA), the enzyme (generally horseradish peroxidase or alkaline phosphatase) is labeled on the antigen or antibody, due to the specific reaction of the antigen and antibody and the enzyme catalyzed substrate Combined with the function, through its color change detection and quantification of a small amount of protein test method, its detection limit (LOD) is 0.1ng to 1μg / mL, widely used in many fields such as food, industry, environmental monitoring, and clinical medicine. Due to the emergence of more and more protein markers in recent years, the requirements for disease detection are also increasing. However, there are several problems...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N33/53
CPCG01N27/447G01N33/53
Inventor 申鹤云张芳芳
Owner BEIJING UNIV OF CHEM TECH
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