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Double signal amplification ELISA detection method based on nanometer gold and graphene oxide

A kind of nano-gold and oxide stone technology, which is applied in measurement devices, biological tests, material inspection products, etc., can solve the problems that there are no nano-gold and graphene oxide, few reports in the field of life science, etc., and achieves strong universal applicability , easy to achieve, good repeatability

Inactive Publication Date: 2014-07-09
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Graphene oxide has only been paid attention to in recent years, and it has been widely used in the field of materials science, but its application in the field of life sciences has rarely been reported.
[0005] At present, there is no report on the simultaneous use of gold nanoparticles and graphene oxide in ELISA detection

Method used

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  • Double signal amplification ELISA detection method based on nanometer gold and graphene oxide
  • Double signal amplification ELISA detection method based on nanometer gold and graphene oxide
  • Double signal amplification ELISA detection method based on nanometer gold and graphene oxide

Examples

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preparation example Construction

[0041] (1) Preparation of gold nanoparticles (AuNPs)

[0042] First, soak the used utensils with aqua regia (mixture of concentrated hydrochloric acid with a mass fraction of 31% and concentrated nitric acid with a mass fraction of 98% at a volume ratio of 3:1), then rinse with double distilled water, and dry for use.

[0043] Add 1 mL of chloroauric acid aqueous solution with a concentration of 1% (mass percent) into 100ml of double-distilled water, stir vigorously and heat to boiling, quickly add 4.5mL of a concentration of 1% (mass percent) sodium citrate aqueous solution, and stir thoroughly , keep boiling for 10min. During this period, the color of the solution changed from gray to blue to purple, and finally to wine red. Remove the heat source, stir for another 15 minutes, and cool naturally to room temperature to obtain gold colloid with an average particle size of 16 nm (characterized by transmission electron microscopy), and store it at 4°C for later use. Transmissi...

Embodiment 1

[0059] Embodiment 1, the method utilizing nano gold and graphene oxide to carry out ELISA detection

[0060] Materials and reagents used in the double signal amplification ELISA detection based on nano gold and graphene oxide of the present invention are as follows:

[0061] (1) Enzyme plate.

[0062] (2) Coating agent: HSP70-Ab1 (mouse IgG).

[0063] (3) Substance to be tested: HSP70 protein standard.

[0064] (4) Primary antibody labeled with gold nanoparticles: the primary antibody labeled with gold nanoparticles is the primary antibody (HSP70-Ab1 (rabbit IgG)) that specifically binds to the substance to be tested (HSP70 protein) and the primary antibody that does not bind to the substance to be tested (HSP70 protein) Mixture of specific binding auxiliary primary antibody (GST-Ab1) (mass ratio 2:3).

[0065] (5) Graphene oxide-labeled enzyme-labeled secondary antibody: The secondary antibody labeled with graphene oxide is HRP-GSTAb2.

[0066] (6) Chromogenic solution: T...

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Abstract

The present invention discloses a new use of nanometer gold and graphene oxide, wherein the new use is specifically an application in ELISA detection signal amplification. The present invention further discloses an ELISA detection method, which comprises: (1) adding an analyte capable of specifically binding with coating antigen to an enzyme label plate loaded with the coating antigen; (2) adding a nanometer gold marker to the enzyme label plate, wherein the nanometer gold marker is a substance obtained by concurrently binding nanometer gold, a protection A (capable of specifically binding with the analyte) and a protein B (incapable of binding with the analyte); and (3) adding a graphene oxide marker to the enzyme label plate, wherein the graphene oxide marker is a substance obtained by binding graphene oxide and a protein C (concurrently binding with the protein A and the protein B). According to the present invention, the nanometer gold and the graphene oxide are concurrently introduced into the conventional ELISA method, such that the double signal amplification is achieved, the detected protein signal is amplified by more than 10 times, and the cost is substantially reduced.

Description

technical field [0001] The invention relates to a new application of nano-gold and graphene oxide, in particular to an application of nano-gold and graphene oxide in amplifying ELISA detection signals. Background technique [0002] Enzyme-Linked Immunosorbent Assay (Enzyme-Linked Immunosorbent Assay, ELISA) is a commonly used analysis and detection method in various biological and chemical laboratories and clinical testing. The basis of ELISA is the solid phase of antigen or antibody and enzyme labeling of antigen or antibody. The antigen or antibody combined on the surface of the solid phase carrier still maintains its immunological activity, and the enzyme-labeled antigen or antibody retains both its immunological activity and enzyme activity. During the determination, the test specimen (the antibody or antigen in it) reacts with the antigen or antibody on the surface of the solid phase carrier. The antigen-antibody complex formed on the solid phase carrier is separated ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6803G01N33/54346
Inventor 钱小红张养军林虹君
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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