Probe for pathological diagnosis of tumor, and preparation method and application thereof

A pathological diagnosis and tumor technology, applied in the field of molecular biology, can solve the problems of peroxidase variability, inconvenient storage and use, and false negative test results.

Active Publication Date: 2013-08-28
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since these methods use natural peroxidase and organic fluorescent dyes, natural peroxidase is easily denatured and inactivated, and it is difficult to purify, expensive, inconveni...

Method used

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  • Probe for pathological diagnosis of tumor, and preparation method and application thereof
  • Probe for pathological diagnosis of tumor, and preparation method and application thereof
  • Probe for pathological diagnosis of tumor, and preparation method and application thereof

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preparation example Construction

[0039] This embodiment also provides a method for preparing a probe for tumor pathological diagnosis, such as figure 1 shown, including the following steps:

[0040] Step S110, mixing the chloroauric acid solution with a concentration of 1-100 mmol / L and the bovine serum albumin or human serum albumin solution with a concentration of 1-500 mg / mL evenly, and then adding a NaOH solution with a concentration of 0.1-10 mol / L , and incubated in a shaker at 0-100° C. for 3-100 hours to obtain gold nanoclusters, wherein the molar ratio of NaOH to chloroauric acid is 1:1-1:100.

[0041] In this embodiment, bovine serum albumin is preferred to prepare gold nanoclusters, because bovine serum albumin has the advantages of non-toxicity, good biocompatibility, and relatively cheap price.

[0042] Step S120 , mixing the gold nanoclusters with the azide reagent pretreated by NHS at a molar ratio of 1:1 to 1:100 to obtain gold nanoclusters with azide groups modified on the surface.

[0043]...

Embodiment 1

[0064] The antibody is an anti-HER2 protein antibody, and the paraffin tumor tissue section is human breast cancer cell MCF7

[0065] 1. Preparation of gold nanocluster probes

[0066] 1. Preparation of gold nanoclusters: at 37°C, add 0.5mL of chloroauric acid solution with a concentration of 50mmol / L to 0.5mL of BSA solution with a concentration of 200mg / mL and mix well; then add 0.2mL with a concentration of 1mol / L of NaOH solution to obtain a mixed solution; the mixed solution was placed in a shaker and incubated at 37°C for 10 hours to obtain gold nanoclusters.

[0067] 2. Azido group modification of gold nanoclusters: mix the above gold nanoclusters with the azide reagent at a molar ratio of 1:1 to obtain gold nanoclusters with azide groups on the surface, wherein the azide reagent Pre-treated by the NHS.

[0068] 3. Alkynyl modification of anti-HER2 protein antibody: Add DBCO-sulfo-NHS Ester alkynylation reagent to anti-HER2 protein antibody and react for 30 minutes, ...

Embodiment 2

[0091] The antibody is an anti-HER2 protein antibody, and the paraffin tumor tissue section is human liver cancer cell HepG2.

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Abstract

The invention relates to a probe for pathological diagnosis of a tumor. The probe comprises a gold nano-cluster and an antibody to a tumor specific marker, wherein the gold nano-cluster and the antibody to the tumor specific marker are connected through a click chemistry manner, and a mol ratio of the gold nano-cluster to the antibody is 1:1 to 1: 100. The invention further provides a preparation method for the probe used for pathological diagnosis of the tumor and a kit including the probe. Since the gold nano-cluster has high peroxide mimic enzyme activity, the gold nano-cluster can catalyze reaction of a color developing agent and emit light in a visible light area, thereby realizing visible light positioning of tumor cells; since the gold nano-cluster has near infrared fluorescence characteristics, the gold nano-cluster emits fluorescent light under the action of exciting light, thereby realizing fluorescence localization of the tumor cells. Thus, the probe for pathological diagnosis of the tumor can realize dual positioning of the tumor cells, so the probe has high accuracy in pathological diagnosis of the tumor.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a probe for tumor pathological diagnosis and its preparation method and application. Background technique [0002] In the past two decades, the mortality rate of cancer has risen sharply. Among middle-aged and middle-aged people aged 35 to 59, tumors have ranked first among various causes of death. Statistics show that the incidence of cancer in my country is about 2 million per 100,000 people, with more than 2.2 million new cases each year and more than 6 million patients being treated. Tumor has become a major cause of the loss of the best labor force and the rise of medical expenses in our country. Although some chemical drugs to prevent tumors have been developed, there is still no primary preventive measure to effectively control tumors. Therefore, early diagnosis is still a basic strategy for tumor prevention and treatment at present and for a long time to come...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574
Inventor 蔡林涛胡德红盛宗海方胜涛张鹏飞王亚楠高笃阳龚萍
Owner SHENZHEN INST OF ADVANCED TECH
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