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Novel functionalized gold nanorod immune probe as well as preparation method and application of gold nanorod biological chip

A technology of gold nanorods and biochips, which is applied in the field of preparation of functionalized gold nanorods immune probes and gold nanorods biochips, and can solve the problem of hindering the coupling of gold nanorods and biomolecules, interfering with biological processes, and unstable connections etc. to achieve stable results, simple operation and high possibility

Inactive Publication Date: 2014-02-12
ZHENJIANG NO 1 PEOPLES HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Some domestic scholars have constructed functionalized gold nanorod immune probes through the electrostatic interaction between gold nanorods and antibodies, but the connection between gold nanorods and antibodies through electrostatic interaction is unstable, and the CTAB in gold nanorod solution Highly toxic, interferes with biological processes and hinders coupling of gold nanorods to biomolecules
However, there has been no report on the construction of gold nanorod immunoprobes by modifying antibodies directly to gold nanorods through strong gold-sulfur bonds through Traut’s reagent.

Method used

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  • Novel functionalized gold nanorod immune probe as well as preparation method and application of gold nanorod biological chip
  • Novel functionalized gold nanorod immune probe as well as preparation method and application of gold nanorod biological chip
  • Novel functionalized gold nanorod immune probe as well as preparation method and application of gold nanorod biological chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Covalent binding of gold nanorods to human IgG antibodies

[0046] (1) Preparation of gold nanorods with a certain aspect ratio:

[0047] Take 0.625mL2mmol / L of chloroauric acid (HAuCl 4 ) solution was added to 1.88mL0.2mol / L CTAB solution, and then 0.45mL0.01mol / L frozen NaBH 4 , and mixed well to obtain a seed solution, which was left to stand in a water bath at 28° C. for 3 hours before use. Take 11.875mL0.2mol / L CTAB solution, add 0.35mL0.01mol / L AgNO 3 solution, after mixing, add 5mL of 0.002mol / L HAuCl 4 , then add 0.16mL of 0.01moI / L ascorbic acid, and finally add 108μL of seed solution, mix well, let it stand in a water bath at 28°C for 1d, centrifuge at 10,000r / min for 25min twice, discard the upper clear night, and weigh the gold nanorods in the lower layer Suspend in 2‐3mL double distilled water and save for later use. The aspect ratio of gold nanorods is about 4:1 ( figure 1 ), uniform in size, good dispersion in aqueous solution, and no agg...

Embodiment 2

[0052] Example 2 Covalent binding of gold nanorods immobilized on glass slides to human IgG antibodies containing fluorescence (FITC)

[0053] (1) Preparation of gold nanorods with a certain aspect ratio: the same as in Example 1.

[0054] (2) Preparation of gold nanorods immobilized on glass slides:

[0055] Take a 7mm×50mm×0.7mm glass slide and wash it with sulfuric acid / hydrogen peroxide (H 2 SO 4 / H 2 o 2 ;3:1v / v) after treatment for 40min, immerse in 10% MPTMS prepared by ethanol at room temperature for 2h to form a mercaptolated silane monolayer on the glass surface, and immerse the treated glass slide in 1mL gold nanorods containing 5mM NaCl 20min, fix the gold nanorods on the glass slide to form a biochip;

[0056] (3) Modification of FITC-containing human IgG antibody with Truat's reagent: the same as in Example 1.

[0057] (4) Functional preparation of gold nanorod biochip:

[0058] Immerse the biochip immobilized with gold nanorods in the human IgG solution t...

Embodiment 3

[0059] Example 3 Detection of gold nanorod biochip immune probe

[0060] (1) Preparation of gold nanorods with a certain aspect ratio: the same as in Experimental Example 1.

[0061] (2) Preparation of gold nanorods immobilized on a glass slide: same as Experimental Example 2.

[0062] (3) Modification of human IgG antibody with Truat's reagent: the same as in Example 1.

[0063] (4) Functional preparation of gold nanorod biochip: same as Experimental Example 2.

[0064] (5) Detection of gold nanorod biochip immune probe

[0065] Add the above-mentioned functionalized gold nanorod biochip immunoprobe into 10nM, 20nM, 40nM, 60nM, 80nM human IgG solution, incubate at room temperature, incubate for 1h at each concentration, wash the unbound IgG with water, read the detection The shift of the longitudinal plasmon absorption peak of the gold nanorod immunoprobe was qualitatively and quantitatively analyzed. The absorption spectrum for detecting human IgG antigen is shown in F...

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Abstract

The invention discloses a novel functionalized gold nanorod immune probe as well as a preparation method and an application of a gold nanorod biological chip. The preparation method comprises the following steps of modifying a human IgG (Intravenous Gamma Globulin) antibody by a Traut reagent; coupling the modified IgG antibody with a sulfydryl having high affinity with gold; and covalently binding the modified IgG antibody with a gold nanorod directly through -SH. The operation method is simple, the conditions are mild, the functionalization of the gold nanorod can be quickly realized, the functionalized gold nanorod fixed on a slide can be used as an unmarked biological chip for specifically detecting a human IgG antigen which can be obtained according to an offset dose-effect relationship curve of a nanorod vertical plasma absorption peak, and each nano vertical plasma absorption peak offset is capable of detecting the human IgG antigen at 137 pM. Therefore, the gold nanorod immune probe prepared by the method and used for antigen-antibody detection has the advantages of simple operation, high detection sensitivity, good specificity and a small number of needed instruments and equipment and is clinically popularized possibly.

Description

technical field [0001] The invention belongs to the technical field of nanometer immunity, and relates to a preparation method and application of a novel functionalized gold nanorod immune probe and a gold nanorod biochip. Background technique [0002] The unique optical properties of gold nanorods (GNRs) make them useful for photothermal therapy, biosensors, molecular imaging, drug delivery for tumors and infectious diseases, etc. In particular, the gold nanorod immunoprobe, compared with traditional detection methods, has outstanding advantages such as no need to label samples, real-time dynamic detection, and high sensitivity. Therefore, it has great advantages in the fields of biomolecular reaction research, immunology, early cancer detection and so on. [0003] But all of these applications require the functionalization of gold nanorods to attach different biochemical groups. However, the functionalization of gold nanorods is a challenging process that usually require...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531
CPCG01N33/54353G01N33/6854
Inventor 汪雪峰唐亮王钧钱伟倪鸿昌
Owner ZHENJIANG NO 1 PEOPLES HOSPITAL
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