Preparation method and application of magnetic sandwich nano immunosensor

An immune sensor and nanotechnology, applied in the direction of instruments, scientific instruments, measuring devices, etc., can solve the problems of uncontrollable antibody orientation, low sensor sensitivity, antibody loss of activity, etc., achieve good immobilization effect, improve sensitivity, and stabilize detection performance Effect

Inactive Publication Date: 2012-10-03
NINGBO UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Among them, the use of physical adsorption method to immobilize antibodies has poor repeatability and low sensor sensitivity; chemical cross-linking and sol-gel embedding cannot control the orientation of antibodies, and it is easy to cause the loss of antibody activity. Therefore, the existing amperometric immunosensors have slow detection speed, Disadvantages of large detection error and low sensitivity

Method used

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  • Preparation method and application of magnetic sandwich nano immunosensor
  • Preparation method and application of magnetic sandwich nano immunosensor

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Embodiment Construction

[0030] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

1. Specific examples

[0031] A method for preparing a magnetic sandwich nano-immunosensor of the present invention specifically comprises the following steps:

[0032] (1) Preparation of secondary antibody probe

[0033] a.Fe 3 o 4 / Preparation of Au gel nano magnetic beads

[0034] a1. 100mL of 0.01wt% chloroauric acid (HAuCl 4 ) placed in a beaker to heat, after boiling, quickly add 2.5mL of 1wt% trisodium citrate, continue to boil for 10min until the solution turns dark red, stir slightly and cool to obtain a colloidal gold solution;

[0035] a2. Add 0.2g of ferric oxide into 90mL of 50wt% ethanol solution and ultrasonically disperse evenly, then add 300uL of 3-mercaptopropyltriethoxysilane (3-MPTES) and 500uL of ammonia water, in a constant temperature water bath at 60°C, Pass N 2 After protection and reaction for 6 hours, wash wit...

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Abstract

The invention discloses a preparation method of a magnetic sandwich nano immunosensor and application of the preparation method. The preparation method is characterized by comprising the following steps: mixing and stirring a collaurum solution and silane ferriferrous oxide into a colorless and transparent solution, and carrying out magnetic separation by externally applying a magnet to obtain FE3O4/Au colloid nano magnetic beads; loading horse radish peroxidase (HRP) and secondary antibodies of an object to be measured to the FE3O4/Au colloid nano magnetic beads to obtain a secondary antibody probe Fe3O4/Au-HRP-Ab2; electrically depositing Au to a glassy carbon electrode, loading primary antibodies of the object to be measured, and then closing non-specific active sites by using protein to obtain an Abl/Au/GCE electrode; and finally, loading antigen to the Abl/Au/GCE electrode, and then obtaining the sandwich nano immunosensor by combining the magnet probe. The magnetic sandwich nano immunosensor can be used for measuring the concentration of melamine, tonyred, clenbuterol or estrogen in food and has the advantages of high detection speed, high accuracy and sensitivity and low cost.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a preparation method and application of a magnetic sandwich nano-immunosensor. Background technique [0002] The amperometric immunoassay method combines high-sensitivity electrochemical sensing technology with specific immune response, and monitors the antigen-antibody reaction through current changes. Accurate results and other characteristics, it is more and more widely used in various fields of society. Its measurement generally includes two steps: first, the enzyme-labeled antibody is bound to the sensor surface through a competitive or sandwich immunoreaction (biological recognition components, such as antibody coating membrane), and then through an enzyme-catalyzed reaction (such as HRP enzyme for H 2 o 2 Catalysis) causes the electrical signal of the test system to change and is measured. Amperometric immunosensors use antigens and antibodies as molecular recognitio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/553
Inventor 胡富陶干宁熊萍朱珊珊
Owner NINGBO UNIV
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